Archives of Microbiology (2021) 203:5849–5857

https://doi.org/10.1007/s00203-021-02575-1

MINI-REVIEW

Recent advances in the research of milbemycin biosynthesis

and regulation as well as strategies for strain improvement
Yu‑Si Yan1 · Hai‑Yang Xia1

Received: 11 May 2021 / Revised: 7 September 2021 / Accepted: 12 September 2021 / Published online: 22 September 2021
© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2021

Abstract
Milbemycins, a group of 16-membered macrocylic lactones with excellent acaricidal, insecticidal and anthelmintic activi-
ties, can be produced by several Streptomyces species. For the reason that they have low toxicity in mammals, milbemycins
and their derivatives are widely used in agricultural, medical and veterinary industries. Streptomyces bingchenggensis, one
of milbemycin-producing strains, has been sequenced and intensively investigated in the past decades. In this mini-review,
we comprehensively revisit the progress that has been made in research efforts to elucidate the biosynthetic pathways and
regulatory networks for the cellular production of milbemycins. The advances in the development of production strains for
milbemycin and its derivatives are discussed along the strain-generation technical approaches of random mutagenesis, meta-
bolic engineering and combinatorial biosynthesis. The research progress made so far indicates that strain improvement and
generation of novel milbemycin derivatives will greatly benefit from future development of enabling technologies and deeper
understanding of the fundamentals of biosynthesis of milbemycin and the regulation of its production in S. bingchenggensis.
This mini-review also proposes that the overproduction of milbemycins could be greatly enhanced by genome minimization,
systematical metabolic engineering and synthetic biology approaches in the future.

Keywords Milbemycins · Regulatory network · Streptomyces bingchenggensis · Strain improvement · Biosynthetic gene
cluster

Introduction environmental profile, as well as unique mode of action on

the insect nervous system, milbemycins and their deriva-
Milbemycins, a group of 16-membered macrolide antibi- tives have been developed as acaricides, insecticides and
otics, can be produced by several Streptomyces species, anthelmintics that are widely applied in agricultural, vet-
including Streptomyces hygroscopicus, Streptomyces gri- erinary, and medical fields (Danaher et al. 2012; Jacobs and
seochromogenes, Streptomyces cyaneogriseus, Strepto- Scholtz 2015). The most active components of milbemycin
myces nanchanggensis and Streptomyces bingchenggensis compounds are milbemycin A3 and A4 (Fig. 1). Milbemec-
(Baker et al. 1996; Carter et al. 1988; Ono et al. 1983a; tin, a mixture of milbemycin A3 and A4, was first com-
Wang et al. 2013, 2020). Milbemycins are structurally and mercialized as an acaricide to control mites circa 1990 in
chemically related to the well-known avermectins produced developed countries (Nicastro et al. 2011; Pluschkell et al.
by Streptomyces avermitilis (Chen et al. 2016). On account 1999). Notably, milbemectin has been used as an effective
of their broad spectrum of activity against various insects insecticide against some insects resistant to avermectins and
and parasites, low mammalian toxicity, a highly favorable their derivatives (Romero-Rodríguez et al. 2015). Nowadays,
a series of derivates from milbemycin A3 and A4 have been
commercialized (Zhao et al. 2011). Milbemycin oxime,
Communicated by Erko Stackebrandt. semi-synthetic analogs of milbemycin A3 and A4, have been
marketed as new veterinary drugs against parasites of pet
* Hai‑Yang Xia
hyxia@tzc.edu.cn animals in Japan, USA, Europe and China (Bienhoff et al.
2013). Additionally, other derivates of milbemycin A3 and
1
Institute of Biopharmaceuticals, Taizhou University, 1139 A4, such as lepimectin and latidectin, have been developed
Shifu Avenue, Jiaojiang District, Taizhou 318000, Zhejiang, as agrochemical products (Kim et al. 2016).
People’s Republic of China

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Fig. 1  Chemical structures of milbemycin A3/A4 and milbemycin family derivatives (color figure online)

Recently, the biosynthetic gene cluster (BGC) of milbe- and regulation and of the various strategies that have been
mycin has been identified in S. bingchenggensis and S. nan- developed to enhance the yields of milbemycins in produc-
changensis NS3226 (Kim et al. 2017; Wang et al. 2010b). tion strains.
The milbemycin BGC consists of ten genes encoding
four multifunctional modular type I polyketide synthases
(milA1, milA2, milA3 and milA4), four tailoring enzymes Milbemycin biosynthesis and regulation
(milC, milD, milE and milF), one regulator (milR) and one
unknown protein (orf1), respectively (Fig. 2A). However, The biosynthesis of milbemycin was first investigated in
unlike the closely situated avermectin BGC, milbemycin Streptomyces hygroscopicus subsp. aureolacrimosus Au-3
BGC is separated by a large DNA fragment (62-kb in S. with 13C isotope labeling in 1983. The carbon skeleton of
bingchenggensis and 55-kb in S. nanchangensis NS3226) milbemycin is derived from seven acetate units and five pro-
between milR and milA1, which is not involved in milbemy- pionate units. The C-25 groups are derived from acetate, pro-
cin biosynthesis. pionate and isobutyrate or dl-valine, respectively. The meth-
Presently, S. bingchenggensis has been developed as a oxyl group at C-5 is from l-methionine (Ono et al. 1983b).
milbemycin producer in Zhejiang Hisun Pharmaceutical Co., It is owing to the structural similarity between milbemycin
Ltd (Li et al. 2017a, b). But the yield of milbemycins in S. and avermectin or meilingmycin that the biosynthetic path-
bingchenggensis is much lower than that of avermectins in way of milbemycin has been proposed based on that of the
S. avermitilis, which hinders the widespread applications of latter (He et al. 2010; Nonaka et al. 1999a, b), namely: (1)
milbemycins in agricultural, veterinary, and medical fields. the formation of starter units acetyl-CoA or propionyl-CoA;
Therefore, there is great potential for commercial reward (2) the elongation of polyketide chain with seven malonyl-
to be realized from improvements in milbemycin overpro- CoA and five methylmalonyl-CoA to starter units by four
duction in Streptomyces species. This mini-review collates giant polyketide synthases (MilA1-MilA4); (3) spiroketal
the current status of research into milbemycin biosynthesis and furan ring formation; (4) post-modification including

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Fig. 2  Biosynthetic gene cluster and pathway of milbemycin in S. biosynthesis; purple, the genes involved in post-modification steps of
bingchenggnesis. A The physical map of milbemycin biosynthetic milbemycin biosynthesis; red, the regulator gene; dash line, the gene
gene cluster. Each arrow indicates the open reading frames (ORFs). with unknown function. B The milbemycin biosynthetic pathway
Green, the PKS genes (milA1, milA2, milA3, milA4) for milbemycin (color figure online)

oxidation, reduction and methylation to form the final milbe- which are precursors of milbemycin A3 and A4 (Wang et al.
mycin components (Fig. 2B). Among the proposed milbe- 2020). Although the function of tailoring enzyme MilC is
mycin biosynthetic pathway, the C5-O-methyltransferase still unknown, the homologs of MilC, MeiC and AveC, have
MilD was determined to catalyze milbemycin A3/A4 to been identified as a spirocyclase that catalyzes the spiroketal
yield B1/B2. The C5-keto reductase MilF was found to be ring formation (Sun et al. 2013).
responsible for reduction of C5-keto groups of milbemycins In recent years, several regulators involved in milbemycin
(Nonaka et al. 2010; Wang et al. 2014; Zhang et al. 2013). biosynthesis have been investigated, but for most of them,
Recently, another tailoring enzyme MilE, has been found to the action mechanisms still have not been clearly elucidated
be involved in furan ring formation in β-family milbemycins (Fig. 3).

Fig. 3  Regulatory network of

milbemycin production in S.
bingchenggnesis. Solid arrow:
activation. Bars: repression.
Solid lines: direct control.
Dashed lines: indirect control
(or unknown) (color figure
online)

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MilR, encoded by a cluster-situated regulatory gene, was hydrolase, a gene family that is not directly involved in
identified as a LAL family (large ATP-binding regulator of milbemycin biosynthesis. The data obtained by Wei et al.
the LuxR family) pathway-specific transcriptional regulator clearly showed that the effect of MilR2 on 5-oxomilbemy-
for milbemycin biosynthesis (Zhang et al. 2016). Deletion cin A3/A4 production might be only partially mediated by
of milR led to the complete loss of milbemycins production. the function of sbi_00791. Additionally, the authors also
Xiang’s group proved that MilR positively regulated milbe- proposed that MilR2 and its homologs probably regulated
mycin biosynthesis by specifically binding the promoters of carbohydrate transport and hydrolysis (Wei et al. 2018).
the milA4-milE operon and of milF with its C-terminal HTH
domain. Four amino acid residues (K37A, K37R, D122A
and D123A) in the N-terminal AAA domain (ATPases Strategies for the enhancement
associated with diverse cellular activity) were shown to be of milbemycin production
essential for the regulatory function of MilR (Zhang et al.
2016). The overexpression of milR significantly improved As broad-spectrum antiparasitic agents, milbemycins and
the production of milbemycin A3/A4, but had no influence their derivatives showed high commercial value and huge
on nanchangmycin biosynthesis in S. bingchenggensis. market potential. Notably, the derivatives from milbemycin
Several other regulators also have been reported to be A3/A4 have higher insecticidal activity and lower toxicity
involved in milbemycin biosynthesis. The pleiotropic regu- than the most commonly used biopesticide—avermectin B ­ 1a.
lator NsdA, a repressor of sporulation and production of However, titer and yield of milbemycins are much lower than
actinorhodin, a calcium-dependent antibiotic and methyl- that of avermectins, which might limit widespread applica-
enomycin, was originally identified in Streptomyces coeli- tions of milbemycins. Scientists have endeavored to improve
color A3(2) (Yu et al. 2006). Previous results showed that the production of milbemycin by process optimization and
the NsdA-encoding gene was widely present and conserved strain improvement. Warr et al. found that concentration of
in Streptomyces species. The disruption of nsdA homologous fructose (or starch) and carbon–nitrogen ratio significantly
gene led to overproduction of milbemycin A4 and nanchang- affected milbemycin production (1994). Subsequently,
mycin in S. bingchenggensis. The nsdA disruption mutant Zhang et al. developed an optimized medium for milbemy-
also showed mycelia pigment and sporulation changes simi- cin production with response surface methodology, which
lar to those of S. coelicolor (Wang et al. 2006). increased the yield of milbemycins to 1110 ± 98 µg/mL, a
γ-Butyrolactone (GBL) systems, including A-factor- 2.61-fold higher yield than before (2011). It is therefore clear
mediated ArpA/AfsA system, SCBs-mediated ScbR/ScbA that improved medium and culture conditions can greatly
system, VBs-mediated BarA/BarX, IM-2-mediated FarA/ enhance strain optimization efforts. The application of dif-
FarX system and SVB1-mediated JadR3/JadW1 system, ferent strategies to improve milbemycin producing strains,
govern secondary metabolites biosynthesis in Streptomy- such as random mutagenesis, metabolic engineering and
ces (O’rourke et al. 2009; Sultan et al. 2016; Takano 2006). combinational biosynthesis, are discussed and summarized
SbbR, a putative ArpA homologous regulator, affected in the following paragraphs.
milbemycin production through directly activating the
expression of milR. SbbA, probably involved in biosynthesis Random mutagenesis
of GBL, had a negative effect on milbemycin biosynthesis
and was repressed by SbbR (Romero-Rodríguez et al. 2015). Random mutation and rational screening is still an efficient
Also, electrophoresis mobility shift assays (EMSAs) proved method to improve productivity of secondary metabolites
that SbbR directly bound to putative promoters of 11 regu- for industrial fermentation (Gao et al. 2010). Wang et al.
latory genes that encode 7 cluster-situated regulators and 4 obtained a high-yield milbemycin-producing strain through
well-known global regulators, which suggests that SbbR/ a rational screening procedure after UV irradiation and
SbbA system, like other ArpA/AfsA-type systems, play an N-methyl-N-nitro-N-nitrosoguanidine (NTG) treatment of
upper-layer role in milbemycin biosynthetic regulatory cas- a parent strain (2009). The production of milbemycin A3/
cades (Romero-Rodríguez et al. 2015). A4 by the final mutant strain reached 1450 mg/L, an 80%
Recently, a novel TetR family regulator MilR2 increase compared to that of the parent S. bingchenggensis
(sbi_00792), tested as an activator for 5-oxomilbemycin strain. Atmospheric and room temperature plasma (ARTP)
A3/A4 biosynthesis, showed little effect on the transcrip- mutation system, new and effective mutation technology,
tion of 5-oxomilbemycin A3/A4 biosynthetic genes in a has also been employed to improve milbemycin yield. For
high 5-oxomilbemycins-producing strain S. hygroscopicus example, Wang et al. obtained a S. bingchenggensis mutant
SIPI-KF (Wei et al. 2018). However, MilR2 was shown to that produced a nearly two-fold higher milbemycin A3/A4
repress the transcription of sbi_00791 by binding to the pro- titer than the initial strain (2014). Ribosomal engineering
moter of sbi_00791. The gene sbi_00791 encodes a putative with random mutagenesis showed great potential to improve

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production of milbemycins. Lu et al. achieved more than production and mildly enhanced the titer of milbemycin A3/
63.3% increasement of milbemycin production by screen- A4 (Wang et al. 2020). MilF transforms 5-oxomilbemycin
ing streptomycin-resistant strain after UV mutagenesis A3/A4 to milbemycin A3/A4 and disruption of milF was
(2018). Teng et al. obtain milbemycin A3 or A4 single found to block the production of milbemycin A3/A4 and B2/
component strain by combinatorial mutagenesis of UV, B3 and resulted in mainly the production of 5-oximilbemy-
ethylmethane sulfonate, atmospheric and room temperature cin A3/A4, which is beneficial for the production of milbe-
plasma (ARTP) and screening strains with glycine-resist- mycin oxime in industry (Wang et al. 2010a).
ance, rifampicin-resistance, sodium acetate-resistance, and Furthermore, enhancement of the utilization of carbon
3-bromopyruvate tolerance. The milbemycin A4 production sources through sugar transporter engineering is also an effi-
of strain 75–22 has increased about four times than that of cient approach for the overproduction of secondary metabo-
initial strain CGMCC 7677 in 50L fermenter. The ratio of lites (Bertram et al. 2004; Getsin et al. 2013; Li et al. 2010).
milbemycin A4 was more than 80%. The milbemycin A3 TP2 and TP5, two ATP-binding transporters, can elevate the
production of strain 95–23 has increased about nine times utilization rate of sucrose in S. bingchenggensis. In the work
than that of initial strain CGMCC 7677 in 50L fermenter. of Jin et al. the expression of TP2 and TP5 was fine-tuned
And the ratio of milbemycin A3 was more than 70% (Teng by native temporal promoters, which resulted in an increase
et al. 2019). of milbemycin A3/A4 production of 36.9% (Jin et al. 2020).
Meanwhile, titers of avermectin B ­ 1a and nemadectin were
Metabolic engineering also greatly improved through controlling the expression of
these two transporters in S. avermitilis and S. cyaneogriseus,
In comparison to the time-intensive and laborious respectively.
approaches of classical mutagenesis highlighted above, Recent years, multi-omics techniques have been devel-
metabolic engineering is regarded as a more efficient and oped as effective ways to identify candidate beneficial muta-
direct method for the improvement of secondary metabolites tions for antibiotics production (Chae et al. 2017; Palazzotto
production in microbes. and Weber 2018). Genome re-sequencing of the high-yield-
Rewiring regulatory network is an efficient approach to ing mutant strain S. bingchenggensis BC04 found 3 candi-
enhance the production of antibiotics and here we highlight date beneficial genes (or operons) (sbi_04868, sbi_06921
the application of this approach in the case of milbemy- and sbi_06922, and sbi_04683) involved in primary metabo-
cin. The overexpression of cluster-situated positive regu- lism from 133 mutation sites. Overexpression of sbi_06921
latory gene milR by Zhang et al. enhanced the production and sbi_06922, and sbi_04683, and downregulation the tran-
of milbemycin A3/A4 by 38% (2016). Wang et al. found scription level of sbi_04868 by CRISPRi system resulted
that the deletion of negative regulatory gene nsdA and sbbA in the titer of milbemycin A3/A4 increasing by 27.6% to
increases milbemycin A3/A4 production by 33.3% and 25%, 3164.5 mg/L (Liu et al. 2021).
respectively (2006). Wei et al. created a milR2-overexpres-
sion strain that produced 34.4% higher levels of 5-oxomilbe- Combinatorial biosynthesis
mycin A3/A4 than the parental strain (2018).
Blocking branches of biosynthetic pathways is generally In the past decades, combinatorial biosynthesis has been
known to result in improvements in the production of sec- widely applied in Type I Polyketide research, particularly
ondary metabolites (Meng et al. 2016; Komatsu et al. 2010; research pertaining to the modular architecture of repetitive
Ikeda et al. 2014). The milbemycin biosynthetic pathway domains in multifunctional Polyketide Synthases (PKSs)
is illustrated in Fig. 2B. MilD (C5-O-methyltransferase) (Baltz 2014; Park and Yoon 2019). An example of com-
introduces methyl group to the C5-OH. There are several binatorial biosynthesis applied in the search for milbemy-
examples of the biosynthetic pathway disruption approach in cin high-yield strains is the work by Kim et al. whereby,
metabolic engineering efforts for the creation of high-yield due to the similarity between milbemycin and avermectin
mutants for the production of milbemycins. For example, the PKSs, a new milbemycin-producing strain was engineered
milD disruption mutant accumulated high milbemycin A3/ based on an avermectin high-producing strain, by replace-
A4 yields through blockage of the generation of C5-O-meth- ment of AveA1 and AveA3 with the MilA1 and MilA3 from
ylmilbemycin B2/B3 (Wang et al. 2014; Zhang et al. 2013). S. bingchenggensis, respectively (Kim et al. 2017). How-
MilE, a CYP171 family cytochrome P450 enzyme, catalyzes ever, the production of milbemycin A3/A4 in the recom-
the conversion of β-family milbemycins to milbemycin A3/ binant strain was only a few micrograms per liter, which
A4. Overexpression of milE increased the titer of milbemy- was much lower than that of wild-type S. bingchenggen-
cin A3/A4 by 53.1% (Wang et al. 2020). CYP41 is respon- sis (Kim et al. 2017). In another case of the application of
sible for the hydroxylation at C26 of milbemycin A3/A4 combinatorial biosynthesis in milbemycin and derivatives
and disruption of cyp41 abolished most milbemycin α9/10 research, production optimization of Tenvermectin A and B

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in S. avermitilis was achieved using a mix of various well- (fermentation), and downstream (recovery and purifica-
established genetic-engineering techniques, protein engi- tion) processes altogether to develop microbial strains with
neering techniques, and synthetic biology techniques. In this desired functions and characteristics (Ko et al. 2020; Rob-
work by Huang et al., Tenvermectin A and B (TVMs), two ertsen et al. 2018). An example that highlights the potential
novel avermectin derivatives, were produced by a geneti- of Systems Metabolic Engineering in the development of
cally engineered strain, S. avermitilis MHJ1011, which was milbemycin producer strains is the work of Liu et al., who
constructed by replacement of the native aveA1 with milA1 obtained a strain with a titer of milbemycin A3/A4 increased
from S. hygroscopocis in S. avermitilis G8-17, an avermec- by 27.6% to 3164.5 mg/L through manipulating three candi-
tin industrial strain. The total titer of the two TVMs was date primary metabolism genes from genome re-sequencing
3400 mg/L in S. avermitilis MHJ1011 (Huang et al. 2015b). results. In essence, this increase in milbemycin yield was
Furthermore, the substitution of aveDH2-KR2 located in achieved by enhancing the uptake of gluconate (2021).
aveA1 of S. avermitilis NA-108 with milDH2-ER2-KR2 Multi-Omics techniques, including genomics, transcrip-
located in milA1 of S. bingchenggensis led to a recombinant tomics, proteomics and metabolomics, have enabled system-
strain which produced ivermectin B1a with a high yield of level analysis of milbemycin biosynthesis and metabolism
3450 mg/L (Zhang et al. 2015). All these efforts have shown in S. bingchenggensis, which will extend the limits of what
that combinatorial biosynthesis is a worthwhile approach in Systems Metabolic Engineering can achieve and offer fur-
the development of high-yield strains for the production of ther opportunities for workers via the identification of rate-
milbemycin and its derivatives. limiting steps in milbemycin metabolism and related path-
ways, which can then be targeted for engineering. Moreover,
the multi-omics data can be used to model the metabolic
Conclusions and future prospects network of milbemycin in S. bingchenggensis, for the predic-
tion of candidate regions for genome minimization.
In past decades, the yields of milbemycins have been Synthetic biology is the third approach we suggest. Syn-
improved a lot through various approaches, such as optimi- thetic biology, as a cutting-edge technology, has shown great
zation of fermentation medium, classical mutagenesis, meta- potential that derives from its robust and reliable charac-
bolic engineering and combinatorial biosynthesis. However, ter. However, the application of synthetic biology greatly
the production yields of milbemycins are still much lower depends on the availability of enabling technology and
than those of avermectin B­ 1a. We propose that the following knowledge, such as the availability of annotated genomes
three strategies would be the most promising for engineering where information about Promoters, Ribosome Binding
milbemycin high-production strains in the future. Sites, Terminators, etc., is accessible. Hence the applica-
Genome minimization (Li et al. 2017a, b) is the first fea- tion of synthetic biology in Streptomyces species, unlike in
sible approach we propose as the next frontier in further Escherichia coli and Saccharomyces were comprehensively
efforts to create milbemycin high-yield production strains. annotated genomes and cell manipulation techniques are
Deletion of competitive gene clusters and non-essential available, is relatively more hindered by the paucity of ena-
chromosomal regions has already been shown to improve bling genetic knowledge and cell manipulation techniques.
yields in the production of avermectin, salinomycin and Nevertheless, there are still ways and means to circumvent
actinorhodin, possibly by enhancing precursor supply these limitations that can allow application of synthetic biol-
(Gomez-Escribano and Bibb 2011; Huang et al. 2015a; Lu ogy to Streptomyces with the goal of creating strains with
et al. 2016). The genome of S. bingchenggensis comprises enhanced production of milbemycin. For example, strains
at least 47 secondary metabolite biosynthetic gene clusters can be designed and built by predictive modelling, refactor-
including that of milbemycin, in which more than 23 are ing biosynthetic pathway, regulatory engineering, product
putative polyketides clusters (Wang et al. 2013). Recently, efflux and metabolic flux manipulation (Breitling et al. 2021;
the Cpf1-based genome editing method has been developed Lee et al. 2019; Phelan et al. 2017). Heterologous expres-
for efficient large fragment deletion in S. bingchenggensis sion or multi-copies integration of secondary metabolic
(Li et al. 2018), which will facilitate the genome minimiza- gene clusters has been successful for actinorhodin, spinosad,
tion in S. bingchenggensis. However, the big downside of rapamycin and pristinamycin (Tan et al. 2017) and could
this approach is that a lot of endeavor is still required to be extended to milbemycin. The recently developed EQCi
decide which areas are candidates for deletion. (Endogenous Quorum-sensing system with CRISPRi) strat-
The second approach we propose is System Metabolic egy has improved rapamycin production by achieving bal-
Engineering. Systems Metabolic Engineering, which inte- ance between essential pathways and product synthesis (Tian
grates traditional metabolic engineering with systems et al. 2020) and could be extended to milbemycin as well.
biology, synthetic biology, and evolutionary engineering, Also, dynamic metabolic control strategies can be helpful
considers upstream (raw material preparation), midstream to decouple the cell growth and product biosynthesis, which

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Archives of Microbiology (2021) 203:5849–5857 5855

can speed up the overproduction of secondary metabolites strain improvement. FEMS Microbiol Lett 368(10):fnab060.
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standing the biosynthesis and regulation of milbemycins, 007
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Acknowledgements We thank the members of Institute of Biophar- Danaher M, Radeck W, Kolar L, Keegan J, Cerkvenik-Flajs V (2012)
maceuticals, Taizhou University for many fruitful discussions. Many Recent developments in the analysis of avermectin and milbe-
thanks to Dr. Alexander Zawaira (formerly of Taizhou University and mycin residues in food safety and the environment. Curr Pharm
now of Beijing Open University, Haidian District, Beijing) for editing Biotehnol 13(6):936–995. https://​doi.​org/​10.​2174/​13892​01128​
and suggestion. 00399​068
Gao H, Liu M, Zhuo Y, Zhou XL, Liu JT, Chen DF, Zhang WQ, Guo
Funding This study was funded the Natural Science Foundation of ZX, Peng S, Zhang LX (2010) Assessing the potential of an
Zhejiang Province to HX (LY19C010002) and the Scientific Research induced-mutation strategy for avermectin overproducers. Appl
Foundation of Taizhou to HX (No. 2001xg07). Environ Microbiol 76(13):4583–4586. https://​doi.​org/​10.​1128/​
AEM.​01682-​09
Declarations Getsin I, Nalbandian GH, Yee DC, Vastermark A, Paparoditis PCG,
Reddy VS, Saier MH Jr (2013) Comparative genomics of trans-
port proteins in developmental bacteria: Myxococcus xanthus and
Conflict of interest All authors declare that they have no conflict of Streptomyces coelicolor. BMC Microbiol 13(1):279–279. https://​
interest. doi.​org/​10.​1186/​1471-​2180-​13-​279
Gomez-Escribano JP, Bibb MJ (2011) Engineering Streptomyces coe-
Ethical statement This manuscript is in compliance with ethical stand- licolor for heterologous expression of secondary metabolite gene
ards. This manuscript does not contain any studies with human partici- clusters. Microb Biotechnol 4(2):207–215. https://​doi.​org/​10.​
pants or animals performed by any of the authors. 1111/j.​1751-​7915.​2010.​00219.x
He YL, Sun YH, Liu TG, Zhou XF, Bai LQ, Deng ZX (2010) Clon-
ing of separate meilingmycin biosynthesis gene clusters by use
of acyltransferase-ketoreductase didomain PCR amplification.
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