polymers

Review
Valorization of Algal Biomass to Produce Microbial
Polyhydroxyalkanoates: Recent Updates, Challenges,
and Perspectives
Anand Narayanasamy 1,† , Sanjay K. S. Patel 2,3,† , Neha Singh 1 , M. V. Rohit 1 and Jung-Kul Lee 3, *

1 Bioconversion Technology Division, Sardar Patel Renewable Energy Research Institute, Vallabh Vidyanagar,
Anand 388120, Gujarat, India; narayanasamy.anand@gmail.com (A.N.); bio4@spreri.org (N.S.);
bio3@spreri.org (M.V.R.)
2 Department of Biotechnology, Hemvati Nandan Bahuguna Garhwal University (A Central University),
Srinagar 246174, Uttarakhand, India; sanjaykspatel@hnbgu.ac.in
3 Department of Chemical Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu,
Seoul 05029, Republic of Korea
* Correspondence: jkrhee@konkuk.ac.kr; Tel.: +82-2-450-3505
† These authors equally contributed to this work.

Abstract: Biopolymers are highly desirable alternatives to petrochemical-based plastics owing to their
biodegradable nature. The production of bioplastics, such as polyhydroxyalkanoates (PHAs), has
been widely reported using various bacterial cultures with substrates ranging from pure to biowaste-
derived sugars. However, large-scale production and economic feasibility are major limiting factors.
Now, using algal biomass for PHA production offers a potential solution to these challenges with
a significant environmental benefit. Algae, with their unique ability to utilize carbon dioxide as a
greenhouse gas (GHG) and wastewater as feed for growth, can produce value-added products in
the process and, thereby, play a crucial role in promoting environmental sustainability. The sugar
recovery efficiency from algal biomass is highly variable depending on pretreatment procedures due
Citation: Narayanasamy, A.; Patel,
to inherent compositional variability among their cell walls. Additionally, the yields, composition,
S.K.S.; Singh, N.; Rohit, M.V.;
Lee, J.-K. Valorization of Algal and properties of synthesized PHA vary significantly among various microbial PHA producers
Biomass to Produce Microbial from algal-derived sugars. Therefore, the microalgal biomass pretreatments and synthesis of PHA
Polyhydroxyalkanoates: Recent copolymers still require considerable investigation to develop an efficient commercial-scale process.
Updates, Challenges, and This review provides an overview of the microbial potential for PHA production from algal biomass
Perspectives. Polymers 2024, 16, 2227. and discusses strategies to enhance PHA production and its properties, focusing on managing GHGs
https://doi.org/10.3390/ and promoting a sustainable future.
polym16152227

Academic Editors: Alberto Romero Keywords: algal biomass; biodegradable plastics; environmental pollution; polyhydroxyalkanoates;
García, Víctor Manuel Pérez Puyana reducing sugars
and Mercedes Jiménez-Rosado

Received: 22 June 2024

Revised: 28 July 2024
1. Introduction
Accepted: 30 July 2024
Published: 5 August 2024 Plastics are polymers widely used in various human activities because of their excep-
tional physicochemical properties, affordability, and practicality [1,2]. However, plastics
pose significant environmental hazards attributed to their slow degradation under normal
circumstances, resulting in their accumulation in nature [3,4]. To address this issue, research
Copyright: © 2024 by the authors. is underway to develop biodegradable plastics [5–7]. Among the different biopolymers
Licensee MDPI, Basel, Switzerland. produced metabolically by various microorganisms, polyhydroxyalkanoates (PHAs) have
This article is an open access article emerged as promising alternatives to plastics [8,9]. PHAs are biodegradable; however,
distributed under the terms and they are produced in only limited amounts by certain prokaryotes and eukaryotes [10].
conditions of the Creative Commons Specific microorganisms can synthesize poly-β-hydroxybutyrates (PHBs), a class of PHAs.
Attribution (CC BY) license (https://
However, the fragile nature and poor physicochemical properties of PHBs limit their scope
creativecommons.org/licenses/by/
for biotechnological applications and, thus, their commercialization [11,12]. Additionally,
4.0/).

Polymers 2024, 16, 2227. https://doi.org/10.3390/polym16152227 https://www.mdpi.com/journal/polymers

Polymers 2024, 16, 2227 2 of 18

the cost of this bioprocess approach is affected by feed costs, which can account for up to
45% of the total cost [9,10]. These restrictions have resulted in a search for biowaste as an
alternative feedstock for PHA production [12,13].
Microbial fermentation of biowaste is more challenging than that of pure sugars be-
cause of its complex nature and diverse composition. Therefore, pretreatment is required
for better accessibility to enzymatic/microbial hydrolysis, facilitating the production of
fermentable sugars or bioactives [14–16]. Biological methods are generally helpful, owing to
the clean transformation of biowaste into biofuels, biopolymers, or other value-added prod-
ucts [9,17–19]. PHAs are aliphatic polyesters produced by numerous prokaryotic organisms,
which can constitute up to 90% of their dry cell weight (DCW) [10]. Fundamentally, the
synthesis of PHAs occurs under physiological stress conditions, including an abundance
of carbon (C) sources in the growth environment and lower concentrations of essential
elements, such as magnesium, potassium, nitrogen (N), phosphorus (P), sulfur, or iron,
during their development period [10,20]. PHAs can be blended into either homopolymer
or copolymer structures. PHB is the most popular homopolymer of PHA and is produced
by many microbes. The biosynthesis of PHB involves three crucial steps: (i) the initial
step is catalyzed by β-ketothiolase (encoded by phaA, which involves the condensation of
two acetyl-CoA molecules to form acetoacetyl-CoA); (ii) the acetoacetyl-CoA is reduced
to acetoacetyl-CoA by NADPH-dependent acetoacetyl-CoA dehydrogenase (encoded by
phaB); (iii) finally, the monomers [(R)- 3-hydroxybutyryl-CoA] are polymerized into PHB by
PHB synthase (encoded by phaC) [9,21]. The type of PHB produced by various organisms
depends on the phaC present in specific organisms [10,11].
Microorganisms, including Gram-positive bacteria such as Bacillus, Rhodococcus, and
Staphylococcus; Gram-negative bacteria such as Acinetobacter, Azotobactor, Burkholderia,
Halomonas, Klebsiella, Pseudomonas, and Ralstonia; algae such as Arthrospira, Botryococcus,
Chlamydomonas, Chlorella, Nostoc, and Spirulina; and engineered microorganisms such as
Escherichia, Halomonas, Bacillus, and Saccharomyces have been reported for PHA produc-
tion [10,22–24]. Microbial PHA production via fermentation has been widely reported
using—(i) pure sugars such as glucose, fructose, maltose, and starch and (ii) biowastes
of diverse origins, including agricultural (rice straw, wheat straw, potato, onion, carrot,
cauliflower, tomato, pea-shells, orange peels, grape peels, melon, and apple pulp), munic-
ipal (vegetable, fruit, and other food waste), industrial (algal biomass, molasses, cheese
whey, biodiesel waste, and dairy waste), and synthetic (wastewater) sources [9,23,25]. Mi-
crobial PHAs are classified into two significant subdivisions based on the C chain length of
their monomeric units: short-chain-length (scl) PHAs with 3–5 C atoms and medium-chain-
length (mcl) PHAs with 6–18 C atoms [26,27].
Microalgae are photosynthetic microorganisms capable of converting carbon dioxide
(CO2 ) and sunlight into biomass [23,28]. They have faster growth rates and do not compete
with food crops for arable land or freshwater resources. Additionally, microalgae can be
cultivated using waste streams such as CO2 derived from flue gas and wastewater, further
improving the sustainability of the production process [29,30]. In estuarine environments,
benthic macroalgae can account for a significant portion of total primary production, some-
times up to 50% [29]. The chemical composition of biomass, including carbohydrates
(7.8–30.8%), proteins (13.0–65.2%), and lipids (3.2–30.4%), is quite variable in red, brown,
and green algae along with various pigments and secondary metabolites [23,28]. The
simple structure of microalgae, which contains less lignin than other renewable biomass,
can facilitate the bioprocessing of their carbohydrate fraction for producing PHAs. Car-
bohydrate recovery from algal biomass highly depends on pretreatment methods, such
as biological, physical, and chemical methods, which significantly vary among types of
algae due to their diverse composition [31,32]. Algae are widely preferred for the bioreme-
diation of toxic compounds from the wastewater of diverse origins and are also effective
in mitigating greenhouse gases (GHGs) such as CO2 . The production of algal biomass
using wastewater or CO2 for biomass production, followed by the further utilization of
biomass hydrolysate to produce PHAs, presents a sustainable approach to achieving a
Polymers 2024, 16, 2227 3 of 18

circular economy [13,33,34]. Moreover, microbes such as Bacillus, Cupriavidus, Halomonas,

Haloferax, Paracoccus, and Rhodotorula demonstrate high potential for producing PHAs
with yields up to 22.5 g/L from algal-biomass-derived sugars [23,35–37]. The microbial
production yield and composition of PHAs from algal biomass are highly dependent on
factors such as pH, temperature, incubation period, the type of limitation (such as N-
and P-sources), the origin of feed, and the kind of PHA-producing culture [31,36,38–40].
Monomeric forms of PHAs, such as PHB, are widely produced by these microbes using
algal biomass [35,36,38,39]. A few studies have reported the microbial production of PHA
copolymers as poly-3-hydroxybutyrate-co-3-hydroxyvalerate [P(3HB-co-3HV)] from algal-
derived sugars or residues [37,41,42]. Generally, copolymer precursors such as valerate
are required for the microbial synthesis of P(3HB-co-3HV) using pure or lignocellulosic-
biomass-derived sugars [4,43]. In contrast, Halomonas mediterranei DSM 1411 can efficiently
produce P(3HB-co-3HV) copolymers lacking any external precursor addition [38]. Addition-
ally, the properties of the produced PHAs must be suitable for biotechnological applications,
particularly in tissue engineering and drug carrier biomedical applications [3,43]. Largely
produced PHB by microbes using sugars or algal biomass demonstrate low potential for
biomedical applications. Given the broad benefits of PHA copolymers, it is highly rec-
ommended that they be easily synthesized by various methods, such as altering the feed
sources, production conditions, and the use of engineered microbes [23,44–46]. Therefore,
different technologies require the integration of complementary microorganisms and algal
biomass as feed to produce various PHA copolymers with diverse properties. The recent
pandemic has highlighted the need for waste management approaches and technological
advancements for sustainable development. This review aims to evaluate the existing liter-
ature on microbial PHA production from algal-based biomass hydrolysates. Furthermore,
various strategies and challenges have been discussed to improve PHA production and
develop sustainable approaches that offer environmental benefits.

2. Algal Biomass and Their Valorization Approaches

Pure sugars are the preferred substrates for biotransformation applications. However,
their high cost limits large-scale conversion, owing to their low economic feasibility. There-
fore, inexpensive feedstocks such as sugars derived from biowaste(s) are being explored to
develop an efficient biotransformation system on a pilot scale [14,18,47]. The lignocellulosic
composition of biomass, comprising cellulose, hemicellulose, and lignin, varies significantly
depending on the source of biomass, such as plants or algae [48–50]. The high contents
of lignin, up to 22% in woody biomass, result in a less efficient system feedstock due to
the partial release of fermentable sugars in hydrolysate or the production of undesirable
byproducts inhibiting biotransformation approaches or microbial fermentation [9]. Various
pretreatment approaches, such as physical, chemical, and biological approaches, have
been widely studied; however, optimal pretreatment approaches or their combinations
are under consideration to maximize the extraction of sugars from biomass while limiting
inhibitory components, such as phenolics [51–53]. Algal-derived biomass is highly desir-
able for producing fermentable sugars compared to plant-based biomass because of its low
lignin content (of up to 5%) [54]. In general, microalgal biomass is rich in various proteins
(10–47 wt% DCW), starch components (10–20 wt% DCW), amylopectin (80–90 wt% DCW),
cellulose, and lipids (20–50 wt% DCW) [55–57]. The high lignin content in woody biomass,
especially rice straw, wheat straw, and agro-residues, yielded lower PHAs compared to
microalgal biomass. Of the various strategies involved in the pretreatment of microalgal
biomass, acid and enzymatic hydrolysis are the most commonly used saccharification
methods to convert biomass into reducing sugars [58–61]. Other pre-treatment methods,
especially physical pretreatment methods, can aid in extracting intracellular byproducts
from microalgae; however, they cannot be used to produce the reducing sugars required for
PHA production [13,53,62,63]. Primarily, microalgal biomass, including its hydrolysates,
often contains toxic inhibitory compounds such as phenolics, which can significantly re-
duce microbial growth and fermentative product yields, including PHA accumulation [23].
Polymers 2024, 16, x FOR PEER REVIEW 4 of 18
Polymers 2024, 16, 2227 4 of 18

yields, including
Therefore, PHAdetoxification
hydrolysate accumulation [23].necessary
is often Therefore, hydrolysate
before detoxification
fermentation is often
[13,64–66]. The
necessary
bioprocessbefore fermentation
approaches [13,64–66].
involved in The bioprocess
PHA production using algalapproaches involved in
biomass are presented in PHA
production
Figure 1. using algal biomass are presented in Figure 1.

Process flow
Figure1.1.Process
Figure flow for
forthe
themicrobial
microbialconversion of algal
conversion biomass
of algal to PHAs.
biomass to PHAs.
Wang et al. [67] reported achieving a high reducing sugar yield of up to 90% at an
Wangloading
enzyme et al. [67] reported
of cellulase achieving
(11.5 U/mL) and a high reducing
amylase (304 sugar
U/mL)yield from of up to acid
sulfuric 90% at an
enzyme loading of cellulase (11.5 U/mL) and amylase (304 U/mL)
pretreated Chlorella vulgaris JSC-6 biomass (120 g/L). Enzymatic hydrolysis using a combi- from sulfuric acid pre-
treated
nation Chlorella
of cellulase vulgaris JSC-6 yielded
and amylase biomass up(120 g/L).
to 0.44 Enzymatic
g/g, reducing the hydrolysis usingCastro
sugar content. a combina-
et al.
tion of[68] achieved
cellulase anda lower
amylasereducing sugar
yielded upyield
to 0.44(0.17 g/g
g/g, during acid
reducing thehydrolysis of mixed
sugar content. Castro et
microalgal biomass (dominated by Ankistrosdemus, Chlamydomonas,
al. [68] achieved a lower reducing sugar yield (0.17 g/g during acid hydrolysis of mixed Chlorella, Micromonas,
and Scenedesmus)).
microalgal biomassThis hydrolysis
(dominated byprocess is species-specific,
Ankistrosdemus, and the yield
Chlamydomonas, of reducing
Chlorella, Micromonas,
sugars varies significantly depending on the type of microalgal species. Furthermore,
and Scenedesmus)). This hydrolysis process is species-specific, and the yield of reducing
the suitable ultrasound-based pretreatment conditions for Dictyota dichotoma biomass to
sugars varies significantly depending on the type of microalgal species. Furthermore, the
produce reducing sugar was identified as 4.3% (w/v) algal suspension at 40% amplitude
suitable ultrasound-based
(6.78 MJ/kg) for an incubationpretreatment conditions
of 40 min, resulting in afor Dictyota
yield of 0.16dichotoma biomassofto pro-
g of the sugars/g
duce
driedreducing
biomass [69]. sugarNgamsirisomsakul
was identified aset4.3% al. [70](w/v) algal suspension
demonstrated at 40%
the influence amplitude (6.78
of glucoamy-
MJ/kg) for an incubation
lase supplementation of 40 min, resulting
on alkaline-treated Chlorella in sp.
a yield of 0.16 the
to enhance g ofyield
the sugars/g
of reducing of dried
biomass [69]. Ngamsirisomsakul et al. [70] demonstrated the influence of glucoamylase
sugars and observed an 87% increase (0.281 g/g). Here, the optimum pretreatment condi-
tion reported include
supplementation using 20% of biomass
on alkaline-treated pretreated
Chlorella sp. towith 1.5% sulfuric
enhance the yieldacid,
offollowed
reducing bysugars
a 20 min incubation at 117 ◦ C.
and observed an 87% increase (0.281 g/g). Here, the optimum pretreatment condition re-
liquid hot water pretreatment of Scenedesmus sp. prior to enzymatic saccharifica-
portedThe include using 20% of biomass pretreated with 1.5% sulfuric acid, followed by a 20
tion resulted in high glucose recovery under the optimum conditions of a solid-to-liquid
min incubation at 117 °C.
ratio, temperature, and incubation period of 1:13 (w/v), 147 ◦ C, and 40 min, respectively [71].
TheThe liquidofhot
addition water pretreatment
glucoamylase and celluloseof Scenedesmus
increased thesp. prior yield
glucose to enzymatic
to 14 g/Lsaccharifica-
upon
tion
the resulted
conversion inofhigh glucose recoverywith
the oligosaccharides, under the optimum
an overall reducingconditions
concentration of of
a solid-to-liquid
0.20 g/g.
ratio, temperature,
The glucose recovery and
wasincubation period
five-fold higher thanofthat
1:13in(w/v), 147 °C,
the control and 40
(without min, respectively
pretreatment).
[71]. The addition
Bhushan of glucoamylase
et al. [17] used a crude enzymeand from cellulose increased to
Chlorella pyrenoidosa thesolubilize
glucosemicroalgae
yield to 14 g/L
and produce
upon the conversionreducingofsugars, achieving a yield with
the oligosaccharides, of 0.19ang/g biomass
overall using 5%
reducing (v/v) of
concentration of
enzyme concentration. During the enzymatic hydrolysis process, the
0.20 g/g. The glucose recovery was five-fold higher than that in the control (without pre- enzymes cellulase, xy-
lanase, and pectinase break the β-1,4 link between the cellulose and hemicellulose, yielding
treatment). Bhushan et al. [17] used a crude enzyme from Chlorella pyrenoidosa to solubilize
higher sugar concentrations compared to acid hydrolysis. However, the cost ineffectiveness
microalgae and produce reducing sugars, achieving a yield of 0.19 g/g biomass using 5%
of the enzymatic hydrolysis process hinders its large-scale commercialization, as enzymes
(v/v)
are of enzymeand
expensive concentration. Duringdue
cannot be recycled thetoenzymatic hydrolysis
their denaturation process,
following the enzymes
biomass pre- cel-
lulase,
treatmentxylanase, and pectinase
[72]. Moreover, break
the slow the β-1,4
reaction rate link between hydrolysis
of enzymatic the cellulose and hemicellu-
impedes the
lose, yielding higher sugar concentrations compared to acid
scaling up of biological pretreatment methods. The pretreatment of Eucheuma spinosum hydrolysis. However, the cost
ineffectiveness
(red seaweed) with of theacid
enzymatic
(HCl, 0.3hydrolysis
mol/L) forprocess hinders its
60 min followed by large-scale commercializa-
enzymatic hydrolysis
resulted in a high reducing sugar concentration of 21.4 g/L,
tion, as enzymes are expensive and cannot be recycled due to their denaturation equivalent to 0.27 g/g of algal
following
biomass [73]. Furthermore, Nordic microalgae, including Chlorococcum
biomass pretreatment [72]. Moreover, the slow reaction rate of enzymatic hydrolysis im- sp. MC-1, Desmod-
esmus sp. RUC-2, Coelastrum astroideum RW-1, and Chlorella vulgaris 13-1 biomass grown
pedes the scaling up of biological pretreatment methods. The pretreatment of Eucheuma
in wastewater and BG11 media, showed a significant variation in the recovery of total
spinosum (red seaweed) with acid (HCl, 0.3 mol/L) for 60 min followed by enzymatic hy-
drolysis resulted in a high reducing sugar concentration of 21.4 g/L, equivalent to 0.27 g/g
of algal biomass [73]. Furthermore, Nordic microalgae, including Chlorococcum sp. MC-1,
Desmodesmus sp. RUC-2, Coelastrum astroideum RW-1, and Chlorella vulgaris 13-1 biomass
grown in wastewater and BG11 media, showed a significant variation in the recovery of
Polymers 2024, 16, 2227 5 of 18

sugar following pretreatment with acid (H2 SO4 ) and enzymatic hydrolysis [31]. Here, the
maximum sugar production reached up to 16.2 g/L. The enzymes secreted by Aspergillus
niger IB-34 showed high hydrolytic activity towards Chlamydomonas reinhardtii biomass [19],
achieving a nearly complete saccharification of Chlorella sorokiniana and Scenedesmus obliquus
at a biomass loading of 10% (w/v), following mild pretreatment at 80 ◦ C for 10 min. Among
the various treatments of algal biomass, including acidic, enzymatic, and microwave
laser-hydrogen peroxide-Fe-nanoparticle (Mv-H2 O2 -Fe) pretreatments, the Mv-H2 O2 -Fe
pretreatment demonstrated the highest total sugar release of 0.99 g/g of DCW, exceeding
the yields of acid and enzymatic treatments, which were 0.59 and 0.49 g/g of DCW, respec-
tively [74]. Furthermore, the Mv-H2 O2 -Fe-treated algae exhibited a maximum biopolymer
production of 0.74 g/g of DCW. The chemo-enzymatic hydrolysis procedure employed
for various microalgae species, including Tetraselmis striata, Tetraselmis sp., Cylindrotheca
fusiformis, Nanofrustulum sp., Picochlorum maculatum, Phaeodactylum tricomutum, Chlorella
sorokiniana, and Chlamydomonas reinhardtii, resulted in a maximum reducing sugar produc-
tion of 34 g/100 g of biomass [75]. Thus, the recovery of sugars from algal biomass is highly
influenced by pretreatment approaches. Therefore, selective strategies or combinations of
pretreatment methods can be beneficial for the effective valorization of algal biomass for
the production of value-added byproducts through fermentation [76–80].

3. Production of Polyhydroxyalkanoates from Algal-Biomass-Derived Sugars

Diverse groups of microbes (~200), such as Firmicutes (Clostridium and Bacillus) and
Proteobacteria, produce PHAs [10,81]. The production of PHAs significantly varies among
these microbes because of their diverse substrate-utilization efficiencies and the different
physiological conditions required for PHA accumulation. PHAs, particularly PHBs, are
generally produced as a homopolymer. Moreover, PHA producers utilize the following
pathways: (i) the methylmalonyl-CoA pathway, (ii) an all-over-again unsaturated fatty acids
engineered pathway, and (iii) a five-step metabolic pathway aided by two stereospecific
2-enoyl-CoA hydratases preceding polymerization [11]. Algal biomass is a suitable low-
cost feedstock for producing PHAs and other value-added products through microbial
fermentation [82–85].
Khomlaem et al. [38] evaluated the potential of various bacterial cultures for PHA
production, including Bacillus megaterium ALA2, Cupriavidus necator KCTC 2649, and H.
mediterranei DSM 1411, using a Chlorella sp. biomass hydrolysate. Under batch mode
(0.2 L), these bacterial species accumulated PHA in the range of 0.84–7.51 g/L, with content
up to 29–75% of their DCW. The PHA content within their biomass showed remarkable
variation, which can be attributed to factors such as genetic stability, fast growth, and
efficient metabolic activities [38]. Abdelmalek et al. [41] demonstrated the production of
PHA using the marine bacteria Halomonas spp., Halomonas pacifica ASL 10, and Halomonas
salifodiane ASL 11, cultivated on the hydrolysates of Spirulina sp., Corallina mediterranea,
and Pterocladia capillacea. Following an acid pretreatment, the bacteria H. pacifica ASL 10
and H. salifodiane ASL 11 could accumulate PHA up to 67 and 63% of DCW, respectively.
These Halomonas spp. accumulated up to 1.2 g/L of PHA from P. capillacea, 1.5 g/L of
PHA from Spirulina sp., and 3.0 g/L of PHA from C. mediterranea. The higher accumula-
tion of PHA when using C. mediterranea for bacterial growth is primarily attributed to its
generation of higher quantities of glucose during acid hydrolysis compared to other algae.
Hydrolysates from other algae mainly consist of glucuronic and galactose monomers with
polysaccharides containing 4-linked galactose instead of glucose [41]. Bhatia et al. [86]
demonstrated PHA production from the red seaweed Eucheuma spinosum using various
bacterial cultures, including Bacillus, Ralsotonia, and Halomonas. With pure sugars, glucose,
galactose, and their mixture as feed, these cultures produced up to 3.02 g DCW/L biomass.
However, inhibitory components, such as furfural, hydroxymethylfurfural, and acetate,
negatively influenced PHA accumulation in Halomonas sp. YLGW01. Biochar derived from
E. spinosum biomass effectively eliminated up to 88% of phenolics. After phenolic removal,
Halomonas sp. YLGW01 showed a significant increase in PHA production, reaching 3.88 g/L
Polymers 2024, 16, 2227 6 of 18

compared to the control value of 2.58 g/L at 4% NaCl. The PHA content in the biomass
increased up to 61.4% of DCW (6.32 g/L). This finding suggests that high PHA production
feasibility can be exploited using E. spinosum-biomass-derived, inexpensive feed under
non-sterile conditions to develop a sustainable and economically viable production sys-
tem [86]. Wastewater originating from the dairy industry is rich in total sugars, especially
lactose, and can effectively support algal growth. Kusmayadi et al. [35] evaluated the
dairy wastewater feed-based production of C. sorokiniana SU-1 biomass to generate sugar
hydrolysate and co-produce PHA and β-carotene using engineered Rhodotorula glutinis
#100-29. After detoxification, the microalgal hydrolysate yielded a maximum PHA and
β-carotene production of 0.90 and 0.09 g/L, respectively. In addition, scaling up this process
to a 5 L capacity enhanced the production of PHA (1.83 g/L) and β-carotene (0.13 g/L).
Despite the high biomass yield of 11.2 g/L using C. sorokiniana SU-1 biomass hydrolysate,
a low PHA accumulation of 16.3% of DCW was noted, which could be attributed to the
diversion of substrate metabolic flux to other linked metabolisms, i.e., β-carotene as a
co-product [35]. Laminaria japonica biomass was used as a carbon source to produce PHA
using three bacterial isolates: Paracoccus sp. LL1, B. megaterium ALA2, and C. necator
NCIMB 11599 [39]. The algal biomass was hydrolyzed using acids to produce 5.9 g/L
and 6.1 g/L of reducing sugars when treated with sulphuric acid and hydrochloric acid,
respectively. These PHA producers could yield PHA up to 32% of DCW with 2% reducing
sugar concentration. Specifically, C. necator NCIMB 11599 accumulated 1.58 g/L of PHA.
Additionally, in the fed-batch mode of operation of the reactor, the yield of PHA increased
to 49% compared to the batch mode yield of 44% [39].
Dubey and Mishra [40] studied the efficiency of PHA production using halophilic
bacteria grown on glycerol obtained from algal biowaste. Halophilic microbes, including
Halomonas spp., H. daqingensis, and H. ventosae, could grow on microalgal biodiesel waste
residues with 5% NaCl supplementation. These microbes accumulated up to 0.24 g/L (35%
of DCW) of PHA, suggesting that waste biomass can be used efficiently to produce PHA,
offering a cost-effective alternative instead of commercially available reagents for PHA
production [40]. Ghosh et al. [87] used macroalgae as a nonconventional source of sugar to
produce biopolymers. This study explored the growth of H. mediterranei and its production
of PHA using reducing sugars obtained from different types of macroalgal biomass. Green
macroalgae produce more reducing sugars, which were utilized for PHA production.
Specifically, when Ulva sp. was used to grow H. mediterranei, the PHA concentration
reached 2.2 g/L (42% of DCW) [87]. Senko et al. [88] explored the ability of C. necator B8619
to produce PHA from a co-culture of Chlorella sp. and fungal biomass. The mechanical
disruption of the cells and the enzymatic hydrolysis of the biomass yielded a reducing
sugar concentration of approximately 39.4 g/L, and the conversion rate of the sugars to
biopolymers was observed as 0.44 g/L/h. In another study, defatted Chlorella biomass was
pretreated with sulfuric acid and hydrochloric acids to yield approximately 46 and 52 g/L
of reducing sugars, respectively [42]. When Paracoccus sp. LL1 was used to convert the
reducing sugars to PHA, the hydrolysate from the biomass treated with 0.3N HCl yielded
a higher concentration of PHA compared to the other pretreatment procedures. The PHA
concentration reached 1.48 g/L accounting for 37.4% of DCW. In addition, a high content
of carotenoids (6.08 mL/L) was generated as a co-product during PHA production. Upon
scaling up production to 5 L fermenter, a significant increase of 144% (3.62 g/L) and 92%
(11.7 mg/L) in PHA and carotenoid production was observed, respectively. Compared to
commercial glucose as a sugar source, the PHA yield was 92% higher when microalgal
hydrolysate was used to grow Paracoccus sp. LL1 [42]. Algal biomass pretreatment with
acids or alkalis is generally required to produce PHAs via microbial fermentation. Kargupta
et al. [83] demonstrated a greener one-pot method to produce PHA using the bacterium
Saccharophagus degradans without a pretreatment of brown seaweed. Additionally, the
use of a membrane bioreactor resulted in three-fold higher PHA accumulation in batch
cultures using seaweed as feed. Azizi et al. [89] demonstrated that C. nector PTCC1615
could produce PHB up to 54% of DCW with a yield of 3.93 g/L from brown seaweed
Polymers 2024, 16, 2227 7 of 18

(Sargassum sp.) hydrolysates derived using acid pretreatment and enzymatic hydrolysis.
Additionally, the fermentation of red seaweed E. spinosum-derived reducing sugars to
PHA by C. nector CECT4635 resulted in yields of 0.26 g of PHB/g of reducing sugar and a
PHA content of 58% of DCW [73]. Cultures of Chlorococcum spp. MC-1, Desmodesmus sp.
RUC-2, Coelastrum astroideum RW-10, and Chlorella vulgaris 13-1 biomass-derived sugars
resulted in high PHA production by Halomonas halophila, yielding up to 1.04, 0.78, 0.08,
and 0.05 g/L, respectively [31]. Moreover, the maximum PHA content in the biomass
was around 27% of DCW with Chlorococcum sp. MC-1-derived sugars. The lower PHA
production by other algal biomass hydrolysates may be due to the presence of inhibitory
phenolics. However, Nordic-microalgal-derived hydrolysates can be pretreated to detoxify
inhibitory compounds and achieve high PHA production [31].
Under phosphate-limiting and low-dissolved-O2 (5%) conditions, the fed-batch culti-
vation of Halomonas boliviensis DSM15516 using G. corneum hydrolysates resulted in high
PHB accumulation and contents of 21.5 g/L and 41% of DCW, respectively [36]. The
maximum PHA productivity was 0.46 g/L/h. In addition, a low gluconic acid content
(15 g/L) was noted because of the low dissolved O2 . In another study, B. megaterium KCTC
2194 demonstrated a high PHA content of 51.4% of DCW (5.50 g/L) when Gelidium amansii
hydrolysates were used in batch mode [37]. Furthermore, the strategy of an intermittent
feeding of hydrolysate in fed-batch mode enhanced PHA production, resulting in a max-
imum biomass production and PHA accumulation of 10.1 and 5.50 g/L, respectively. In
contrast, the pH-state strategy resulted in a slightly lower biomass production of 8.20 g/L
with a PHA content of 53.2% of DCW [37]. A pilot-scale study (40-L) on PHA production by
H. mediterranei in pneumatically agitated bioreactors (outdoor fermentation) using Ulva sp.
hydrolysate demonstrated high biomass and PHA productivity of 50.1 and 27.0 mg/L/h,
respectively [90]. PHA production reached 56% w/w of biomass with a conversion yield of
0.107 g/g of algal DCW. Using an ultrasonic harvesting approach with energy inputs of
7.8 kWh/m3 resulted in a 30% removal efficiency of H. mediterranei cells [90]. In another
study, the influence of the initial culture density (10–500 g/L) on PHA production by H.
mediterranei using Ulva sp. hydrolysate was evaluated to enhance the conversion yield [91].
Maximum biomass and PHA contents of 56.0 g/L and 49.4%, respectively, were observed
at an initial culture density of 50 g/L. Maximum biomass productivity was 0.05 g/L/h,
and PHA productivity was 0.024 g/L/h. Remarkably, the productivity and PHA pro-
duction were similar to those in the standard media. The average molecular weight was
approximately 920–960 kDa, with a polydispersity index of approximately 1.6. The pri-
mary benefits of using high-cell-density cultivation for PHA production include improved
productivity, reduced downstream processing costs, and effective wastewater treatment.
Economic analysis predicted Ulva sp.’s annual PHA production rate (APPR) cultivated
under offshore conditions would be 148 g PHA/m2 /year. The commercial cost of the PHA
ranged from USD 2.4–5.5 per kg. Based on the APPR, the PHA production from Ulva sp.
could incur an annual income of USD 3369/ha/year [91]. The starch and cellulose extracted
fractions of Ulva sp. by H. mediterranei resulted in lower PHA production yields of 5.1 and
3.5 mg/g dry weight, respectively, compared to the whole biomass yield of 77.8 mg/g
dry weight of Ulva [92]. Furthermore, economic analysis showed that the direct use of
Ulva sp. biomass hydrolysate fermentation for PHA is beneficial. In contrast, glucose and
hydrochar coproduction from Ulva sp. biomass did not provide any remarkable economic
benefits [92]. Novel strains Bacillus pacificus NAA2 and Klebsiella quasipneumonia NAA4
showed a high PHA production of up to 72.7% from seaweed hydrolysate [93]. Details of
the algal-biomass-derived sugars used to produce microbial PHAs are presented in Table 1.
Polymers 2024, 16, 2227 8 of 18

Table 1. Microbial conversion of microalgal-derived biomass hydrolysate into polyhydroxyalkanoates

(PHAs).

Reducing PHA References

Algal Biomass Pretreatment Method Sugar Yield
(g/L) Microbial PHA Producer Composition
(g/L)

Algal biodiesel Halomonas daqingensis 0.24 PHB

-a - [40]
waste Halomonas ventosae 0.21 PHB
Acid and microwave-
Algal biomass 0.99 b Mixed culture 0.74 b PHB [74]
peroxidenanoparticles
Bacillus megaterium ALA2 0.84 PHB
Acid (HCl) 20.0 Cupriavidus necator KCTC 2649 7.51 PHB [38]
Chlorella sp. Haloferax mediterranei DSM 1411 3.79 P(3HB-co-3HV)
Dilute acid (H2 SO4
52.0 Paracoccus sp. LL1 3.62 P(3HB-co-3HV) [42]
and HCl)
Chlorella vulgaris
Acid (H2 SO4 ) 10.8 Halomonas halophila 0.05 PHB [31]
13-1
Acid (H2 SO4 and HCl) and
C. vulgaris C-1 39.4 C. necator B8619 0.44 PHB [88]
mechanical destruction
Chlorella
Acid (H2 SO4 ) 39.8 Rhodotorula glutinis #100-29 1.83 PHB [35]
sorokiniana SU-1
Chlorococcum sp.
Acid (H2 SO4 ) 12.5 H. halophila 1.04 PHB [31]
MC-1
Coelastrum
Acid (H2 SO4 ) 11.4 H. halophila 0.08 PHB [31]
astroideu RW10

Corallina Halomonas pacifica ASL 10 2.80 P(3HB-co-3HV)

Acid (H2 SO4 ) - [41]
mediterranea Halomonas salifodiane ASL 11 3.00 P(3HB-co-3HV)
Desmodesmus sp.
Acid (H2 SO4 ) 16.2 H. halophila 0.78 PHB [31]
RUC-2
Acid (HCl) 21.4 C. necator CECT4635 0.60 b PHB [73]
Eucheuma spinosum
Dilute acid (H2 SO4 ) - Halomonas sp. YLGW01 3.9 PHB [86]
Gelidium amansii Acid (H2 SO4 ) 29.1 Bacillus megaterium KCTC 2194 5.50 P(3HB-co-3HV) [37]
Gelidium corneum Hydrothermal - Halomonas boliviensis DSM15516 21.5 PHB [36]
5.9–6.1 Paracoccus sp. LL1 1.58 P(3HB-co-3HV)
Laminaria japonica Acid (HCl and H2 SO4 ) 5.8–6.0 B. megaterium ALA2 0.65 PHB [39]
5.9–6.1 C. necator NCIMB 11599 2.39 PHB

Pterocladia H. pacifica ASL 10 1.00 P(3HB-co-3HV)

Acid (H2 SO4 ) - [41]
capillacea H. salifodiane ASL 11 1.50 P(3HB-co-3HV)
Sargassum sp. Acid (H2 SO4 and HCl) - C. necator PTCC1615 3.93 PHB [89]
H. pacifica ASL 10 1.30 P(3HB-co-3HV)
Spirulina sp. Acid (H2 SO4 ) - [41]
H. salifodiane ASL 11 1.20 P(3HB-co-3HV)
2.20 P(3HB-co-3HV) [87]
Ulva sp. Subcritical water - H. mediterranei 2.08 P(3HB-co-3HV) [90]
0.12 b P(3HB-co-3HV) [91]
a b
Not reported or available; yield in g/g of weight or g of PHA/g of DCW.

4. Limitations and Challenges in Microbial Production of Polyhydroxyalkanoates from

Algal Biomass
Primarily, sugars used to produce microbial PHAs cover ~50% of the total production
cost; therefore, employing inexpensive and renewable feedstocks such as algal biomass
can reduce production costs [32,91]. Despite several advantages, the commercialization
of microbial PHA production from microalgae-derived sugars has faced numerous chal-
Polymers 2024, 16, 2227 9 of 18

lenges [23,31]. A few disadvantages of using algal biomass are low availability due to the
insignificant number of algal-based industrial sources, lack of desirable sugars or precur-
sors, and restricted seasonal accessibility of macroalgae [28,85]. One of the main challenges
is the high cost of producing microalgae biomass at a large scale, which is often due to
the cost of inputs such as nutrients and energy. Additionally, extracting and refining the
fermentable sugars from algal biomass for microbial PHA production can be complex
and costly [28,31,32]. Still, microbial PHA production is more expensive than petroleum-
based plastic [23,85]. To make microalgae-derived microbial PHAs cost-competitive, a
multi-dimensional approach, such as an algal biorefinery, may be necessary. Largely, PHB
production has been reported by bacteria from algal biomass [31,39,40]. Due to the high
crystalline nature and poor mechanical properties of PHB, its application is highly restricted
over the broad uses of the copolymers of PHA having desirable properties. Still, achieving
the bacterial synthesis of PHA copolymers such as P(3HB-co-3HV) from biomass-derived
sugars requires precursor supplementation such as valerate in the feed [4,43]. Therefore,
the co-digestion of feed containing such precursors and algal biomass can be desirable to
synthesize the corresponding copolymers of PHA by microbes [23,43]. While microalgae
offer a promising alternative to microbial PHA production, significant challenges must be
addressed before their widespread commercialization. The careful consideration of the
cost-effectiveness and scalability of the production process, as well as the optimization of
the extraction and purification steps, will be crucial in overcoming these challenges and
unlocking the full potential of microalgae as a feedstock to produce microbial PHAs [23,32].

5. Mechanical and Physiochemical Properties of the Microbially Synthesized PHAs

from Algal Biomass
As the most common type of PHA production, PHB has been widely reported from al-
gal biomass through microbial fermentation [23,31]. The broader uses of PHB are restricted
because of their stiff and brittle nature with a high degree of crystallinity. Additionally, its
melting temperature (Tm ) and decomposition temperature are nearly close to each other
at 175 and 185 ◦ C, respectively [85]. The properties of PHAs, such as Tm , glass transition
temperature (Tg ), crystallinity, the modulus of elasticity, breaking strengths, and Mw, are
vital for their potential applications. The microbially synthesized P(3HV-co-3HV) as a
copolymer exhibits lower crystallinity than PHB, resulting in its better mechanical prop-
erties, such as Young’s modulus, tensile strength, Tm , and Tg values [23,85]. Therefore,
the high degradation potential of copolymers with improved mechanical and physical
properties proved beneficial over PHB for various applications [23,43]. H. mediterranei and
Ulva sp.-based-synthesized PHAs showed decomposition and Tm of 247 and 176 ◦ C, respec-
tively [91]. Using polystyrene and polymethyl methacrylate standards, the average Mw
and Polydispersity Index (PDI) of synthesized PHAs varied in the range of 0.92–0.96 × 106
and 1.56–1.68, respectively. For the polymer to be of varied usage, the average Mw can
be high with a low heterogeneity (PDI, 1.5–2.0) [91]. In contrast, PHAs produced by H.
daqingensis and H. ventosae from algal biodiesel waste residues showed higher degradation
temperatures of 290–296 ◦ C [40]. The higher degradation temperature can be associated
with the presence of slight impurities. The H. daqingensis-synthesized PHB showed a low
Mw of 0.31 × 106 with a PDI of 1.82 and a Tg of 176 ◦ C [40]. PHA synthesized by enriched
mixed culture from residual algal biomass exhibited Tg and a crystallization temperature
of 298 ◦ C [74]. C. necator PTCC1615 synthesized PHB showed Tm and crystallinity temper-
ature values of 167 and 99.5 ◦ C, corresponding to standard PHB values of 177 and 88 ◦ C,
respectively [89]. The H. mediterranei-based copolymers of PHA from Ulva sp. hydrolysate
exhibited Tm and decomposition temperatures of 150–177 and 241–248 ◦ C under various
synthesis conditions, respectively [87,90]. Therefore, the physiochemical properties of
PHAs, such as thermal stability, mechanical strength, and biodegradability, can be tuned
by varying the bacterial strain, fermentation conditions, and the chemical composition of
the polymer [32,40,90].
Polymers 2024, 16, 2227 10 of 18

6. Genetic Engineering Approaches

PHA production by natural producers is complex owing to the different growth con-
ditions and extended growth periods required to accumulate PHA and its extraction via
hydrolysis from their cells [94–96]. As a commonly used microbe for recombinant studies,
E. coli has been extensively used to study PHA production because of its straightforward
culture and rapid growth [97]. Incorporating PHA synthesis genes into E. coli has enabled
efficient and enhanced PHA production at a lower cost compared to other microorganisms.
However, studies on recombinant microorganisms that produce PHA from microalgal
biomass are limited [80,85]. Developing algal-biomass-based biorefineries presents signif-
icant challenges due to seasonal compositional variations, such as carbohydrate content
and salinity [98,99]. Jiang et al. [100] produced PHB using microalgal biomass as the
substrate. The gene encoding the enzymes involved in the production of PHB by Massila
sp. (isolated from seaweed) was incorporated into E. coli to enhance the PHB yield. A
117% increase in DCW and a 213% increase in PHB production was observed. Sathish
et al. [101] used recombinant E. coli to treat microalgal biomass and produce PHA, resulting
in an accumulation of up to 34% PHB of DCW using pure sugar, which further increased
to 51% of DCW when using microalgal biomass as a carbon source for PHA production.
Rahman et al. [97] reported that genetically modified E. colis secrete PHB directly into
the medium, simplifying the downstream processing and extraction of intracellular PHB
from bacterial biomass. It was observed that the PhaP1 encoding phasin, with a lower
molecular weight, binds to the PHB molecules, thereby reducing the granule size of the
produced PHB and facilitating the secretion of PHB from the cell. In a 5 L fermentation,
the engineered R. glutinis #100-29 exhibited a high production of PHA and β-carotene up
to 1.9 g/L and 4.9 mg/g of reducing sugars from glucose and C. sorokiniana-SU-1-derived
sugars as carbon sources, respectively [35]. Therefore, the co-production of value-added
products during PHA accumulation, such as β-carotene or others, by engineered PHA pro-
ducers, can benefit the economic viability of bioprocesses. Overall, emerging strategies can
prove beneficial for producing PHAs from algal biomass hydrolysates, including designing
metabolic pathways for dynamic control over PHA production, engineering salt-tolerant
halophilic microbes to improve PHA yields, and the synthetic engineering of microbial
consortia for the efficient conversion of diverse algal biomass hydrolysates into desirable
copolymers of PHAs [86,98,99].

7. Biotechnological Applications of Polyhydroxyalkanoates

The bacterial synthesis of PHA using algal biomass and its biotechnological applica-
tions are presented in Figure 2. PHA is a biopolymer that has recently gained significant
attention because of its potential applications as an adsorbent [102,103]. PHAs exhibit
excellent adsorption properties, making them promising materials for various adsorption
applications, such as wastewater treatment, the removal of heavy metals from contaminated
water, and the adsorption of organic pollutants [104,105]. Furthermore, their biodegrad-
ability and renewability render them environmentally friendly alternatives to conventional
adsorbents. The PHA-polyricinoleic acid-Ag nanocomposite exhibited high adsorption
efficiency, achieving up to 99.5% removal of methylene blue from wastewater, drinking
water, and river water samples [103]. The biodegradable PHA-diethanolamine matrix
measured heavy metals such as lead, cadmium, and zinc, with detection limits of 1.05, 0.42,
and 0.13 µg/L in food, wastewater, and water samples, respectively [106]. Additionally,
the PHA-biochar reactor efficiently adsorbed and biodegraded trichloroethylene [102].
PHAs act as potential antioxidants because of their unique properties, which make
them highly effective at scavenging free radicals and protecting cells from oxidative dam-
age [107,108]. Poly-lactic acid/PHA film showed a high total antioxidant activity of 36.0 µg
Trolox equivalent/cm2 , which is helpful in effectively preserving pork meat [109]. A
significant advantage of using PHAs as antioxidants is their biodegradability, an indica-
tion that natural environmental processes can easily break them down, reducing their
impact on ecosystems compared with other synthetic antioxidants [110,111]. Furthermore,
Polymers 2024, 16, 2227 11 of 18

PHAs can be easily synthesized from renewable carbon sources using metabolic engi-
neering and synthetic biology approaches. This makes them promising alternatives to
traditional antioxidants derived from nonrenewable resources, which may harm the envi-
ronment [109,112,113]. Biodegradable nanoparticulate derived from PHA/diethanolamine-
caffeic acid exhibited high antioxidant activity with an inhibitory concentration (IC50 )
of 32.2 mg/L for 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and was
Polymers 2024, 16, x FOR PEER REVIEW 11 of 18
highly degradable in clay soil within 60 days at 25 ◦ C [111]. In another study, bio-based
PHA/tannin films showed remarkable antioxidant activity, achieving 80% scavenging ac-
tivity within 7 h towards DPPH, making them helpful in food-packaging applications [112].
1.05, 0.42, and 0.13
Multifunctional µg/L in food, wastewater,
chromone-incorporated PHA and water
film samples,
exhibited respectively
antioxidant [106].(>55%
activity Ad-
ditionally,
towards DPPH)the PHA-biochar
and superiorreactor efficiently
antimicrobial adsorbed
activity andAspergillus
towards biodegraded trichloroeth-
niger, Escherichia
ylene [102].
coli, and Staphylococcus aureus [114].

Figure
Figure 2.
2. Bacterial conversion of
Bacterial conversion ofmicroalgal
microalgalbiomass
biomasstoto PHAs
PHAs and
and their
their biotechnological
biotechnological applica-
applications.
tions.
Owing to their biocompatibility and bioactivity, these polymers are promising alterna-
tivesPHAs
to conventional antimicrobial
act as potential agents
antioxidants [115]. PHA/diethanolamine-caffeic
because of their unique properties, which acid-based
make
nanoparticles
them exhibit at
highly effective antimicrobial
scavengingactivity, inhibiting
free radicals up to 98.0%
and protecting of food
cells frompathogen
oxidativegrowth
dam-
within 48 h of incubation, including S. aureus DSM683, Listeria monocytogenes
age [107,108]. Poly-lactic acid/PHA film showed a high total antioxidant activity of 36.0 DSM19094,
Salmonella
µg enterica DSM9386,
Trolox equivalent/cm and is
2, which E. helpful
coli DSM787 [111]. Similarly,
in effectively biodegradable
preserving pork meat bacterial-
[109]. A
cellulose-deposited
significant advantageflax fabrics
of using incorporating
PHAs PHA/polylactic
as antioxidants acid showed an
is their biodegradability, prolonged
indica-
antimicrobial
tion that natural activity against E.processes
environmental coli, S. aureus, and Pseudomonas
can easily break them aeruginosa [116]. However,
down, reducing their im-
several
pact on limitations
ecosystemsneed to be considered
compared with otherwhen usingantioxidants
synthetic PHAs for antimicrobial applications:
[110,111]. Furthermore,
(i) their limited spectrum of activity, that is, diverse effectiveness
PHAs can be easily synthesized from renewable carbon sources using metabolic engineer-against Gram-positive and
Gram-negative bacteria and fungi; (ii) the durability of PHAs under
ing and synthetic biology approaches. This makes them promising alternatives to tradi- different environmental
conditions;
tional and (iii)derived
antioxidants the highfrom cost of PHA production
nonrenewable [116,117].
resources, Strategies
which may harmto overcome these
the environ-
limitations have been proposed, such as modifying the PHA
ment [109,112,113]. Biodegradable nanoparticulate derived from PHA/diethanolamine- structure, combining PHAs
with other
caffeic acid antimicrobial
exhibited highagents (nanomaterials),
antioxidant activity withandan optimizing
inhibitoryproduction
concentration processes to
(IC50) of
improve activity, durability, and cost-effectiveness [118].
32.2 mg/L for 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and was highly
PHAs in
degradable have
claygained considerable
soil within 60 daysattention because
at 25 ○C [111]. of their biomedical
In another applications
study, bio-based PHA/tan-and
their biocompatible and biodegradable properties that make them
nin films showed remarkable antioxidant activity, achieving 80% scavenging activity suitable for various med-
ical purposes,
within such as
7 h towards tissuemaking
DPPH, engineering,
them drug delivery
helpful systems, anticancer
in food-packaging activity,
applications and
[112].
surgical implants [119,120]. PHA-g-cellulose-Fe
Multifunctional chromone-incorporated PHA film O /ZnO nanocomposites showed
3 4 exhibited antioxidant activity (>55% an effi-
cient delivery system for artesunate and dopamine drugs with an IC50 of 12.3–21.2 µg/mL,
towards DPPH) and superior antimicrobial activity towards Aspergillus niger, Escherichia
along with robust anticancer activity against HCT-116 cells [118]. Sterilized gauze loaded
coli, and Staphylococcus aureus [114].
with 31.1 µg of AgNPs/gelatin/PHA resulted in an effective wound dressing [120]. PHAs
Owing to their biocompatibility and bioactivity, these polymers are promising alter-
offer advantages, such as biocompatibility, controlled degradation, and the ability to be
natives to conventional antimicrobial agents [115]. PHA/diethanolamine-caffeic acid-
processed into various forms and structures. Their biodegradability ensures that the body
based nanoparticles exhibit antimicrobial activity, inhibiting up to 98.0% of food pathogen
can safely break them down over time, thereby reducing the risk of long-term compli-
growth within 48 h of incubation, including S. aureus DSM683, Listeria monocytogenes
cations [117,120,121]. In addition, the mechanical properties of PHAs can be tailored to
DSM19094, Salmonella enterica DSM9386, and E. coli DSM787 [111]. Similarly, biodegrada-
ble bacterial-cellulose-deposited flax fabrics incorporating PHA/polylactic acid showed
prolonged antimicrobial activity against E. coli, S. aureus, and Pseudomonas aeruginosa
[116]. However, several limitations need to be considered when using PHAs for antimi-
crobial applications: (i) their limited spectrum of activity, that is, diverse effectiveness
against Gram-positive and Gram-negative bacteria and fungi; (ii) the durability of PHAs
Polymers 2024, 16, 2227 12 of 18

meet specific requirements, rendering them versatile for various biomedical applications.
Furthermore, PHAs support cell growth and adhesion, making them ideal for tissue en-
gineering applications [3,122–124]. Researchers have created drug delivery systems with
controlled release capabilities by incorporating bioactive agents such as growth factors
or antibiotics into PHA matrices [117,124]. These characteristics make PHAs promising
materials for various biomedical applications, offering opportunities for advancements in
healthcare and improving patient outcomes [3,120,125].
PHAs have shown promising potential in bioremediation owing to their biodegradabil-
ity and the ability of microorganisms to efficiently degrade various pollutants by utilizing
these polymers as a carbon source [126,127]. The use of PHAs in bioremediation can sig-
nificantly restore polluted environments by providing a sustainable and efficient method
for removing contaminants from soil, water, and air [103,104,127]. Furthermore, PHAs
offer several advantages for bioremediation applications, including their biodegradability,
ability to be utilized as a carbon source and energy reserve by microorganisms, and effec-
tiveness in efficiently degrading various pollutants. Additionally, PHAs can be synthesized
from renewable resources, thus reducing the environmental impact of bioremediation
efforts [3,103,105].

8. Perspectives and Concluding Remarks

Microbial PHAs are biodegradable polymers synthesized by various microorganisms
for storage under nutrient-limiting conditions. However, their production encounters sev-
eral challenges: (i) the limited availability of suitable carbon sources for microbial growth
and PHA production, (ii) the complexity of regulating and controlling the metabolic path-
ways involved in PHA synthesis, (iii) the low yield and productivity of PHA production
in microbial systems, (iv) the challenges associated with scaling up the production of mi-
crobial PHAs to an industrial scale, (v) the need for extensive downstream processing and
purification of microbial PHAs, (vi) the potential risk for contamination in microbial PHA
production processes, and (vii) the challenges of achieving consistent and reproducible
product quality. These challenges hinder the widespread adoption and commercialization
of microbial PHAs as sustainable alternatives to traditional petroleum-based plastics. PHAs
are highly desirable over homopolymers (PHBs) because of their broad biotechnological
applications, especially in the biomedical sector. Generally, precursor supplementation is
required during microbial fermentation to produce the corresponding PHA copolymers.
Therefore, co-substrate strategies, such as the co-digestion of feed, can be employed to
produce desirable copolymers of PHAs with desired variations in their properties. Addi-
tionally, the successful conversion of algal biomass into valuable sugars, including PHAs,
for various biotransformation applications, is highly dependent on effective pretreatment
approaches. Genetically engineered algal cultures show promise for an efficient bioremedi-
ation of contaminated wastewater, or GHGs, such as CO2 utilization (flue gas as origin),
enhancing cell biomass for use as a feedstock for various biotechnological applications,
including PHA production. Utilizing algal biomass as a carbon source to produce microbial
PHAs on a pilot scale offers a sustainable solution for addressing the issue of plastic waste
and reducing dependence on fossil fuels, thereby promoting a circular economy. Further-
more, using genetically engineered PHA for easy recovery or value-added co-product
formation can minimize economic competitiveness compared to plastics. Advances in
molecular and genetic engineering have enabled the selection of bacterial strains capable
of producing PHAs in high quantities, up to 90% of their total biomass. However, the
high cost of production remains a significant challenge for the commercialization of PHAs,
limiting their widespread adoption.
Polymers 2024, 16, 2227 13 of 18

Author Contributions: Conceptualization, A.N., S.K.S.P. and J.-K.L.; data curation, A.N., N.S., M.V.R.
and S.K.S.P.; writing—original draft preparation, A.N. and S.K.S.P.; writing—review and editing,
S.K.S.P. and J.-K.L. All authors have read and agreed to the published version of the manuscript.
Funding: This research was supported by the Basic Science Research Program through the National
Research Foundation of Korea (NRF), funded by the Ministry of Science, ICT, and Future Planning
(NRF-2022M3A9I5015091, 2022M3A9I3082366, and RS-2023-00222078).
Acknowledgments: The research work is financially supported by the Gujarat State Biotechnology
Mission (GSBTM/KD0HB2) and Indian Council of Agricultural Research (ICAR), under the All India
Coordinated Research Project (AICRP) EAAI program, Govt. of India. Also, the authors would like
to thank Vice Chancellor, Hemvati Nandan Bahuguna Garhwal University (A Central University),
Srinagar, Uttarakhand for providing the necessary support.
Conflicts of Interest: The authors declare no conflicts of interest.

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