Microbial Biotechnology最新文献

{"title":"Probiotic Lactobacillus casei Shirota prevents acute liver injury by reshaping the gut microbiota to alleviate excessive inflammation and metabolic disorders","authors":"Ren Yan,&nbsp;Kaicen Wang,&nbsp;Qiangqiang Wang,&nbsp;Huiyong Jiang,&nbsp;Yingfeng Lu,&nbsp;Xiaoxiao Chen,&nbsp;Hua Zhang,&nbsp;Xiaoling Su,&nbsp;Yiling Du,&nbsp;Lifeng Chen,&nbsp;Lanjuan Li,&nbsp;Longxian Lv","doi":"10.1111/1751-7915.13750","DOIUrl":"https://doi.org/10.1111/1751-7915.13750","url":null,"abstract":"<p>Millions of people die from liver diseases annually, and liver failure is one of the three major outcomes of liver disease. The gut microbiota plays a crucial role in liver diseases. This study aimed to explore the effects of <i>Lactobacillus casei</i> strain Shirota (LcS), a probiotics used widely around the world, on acute liver injury (ALI), as well as the underlying mechanism. Sprague Dawley rats were intragastrically administered LcS suspensions or placebo once daily for 7 days before induction of ALI by intraperitoneal injection of D-galactosamine (D-GalN). Histopathological examination and assessments of liver biochemical markers, inflammatory cytokines, and the gut microbiota, metabolome and transcriptome were conducted. Our results showed that pretreatment with LcS reduced hepatic and intestinal damage and reduced the elevation of serum gamma-glutamyltranspeptidase (GGT), total bile acids, IL-5, IL-10, G-CSF and RANTES. The analysis of the gut microbiota, metabolome and transcriptome showed that LcS lowered the ratio of Firmicutes to Bacteroidetes; reduced the enrichment of metabolites such as chenodeoxycholic acid, deoxycholic acid, lithocholic acid, <span>d</span>-talose and <i>N</i>-acetyl-glucosamine, reduce the depletion of <span>d</span>-glucose and <span>l</span>-methionine; and alleviated the downregulation of retinol metabolism and PPAR signalling and the upregulation of the pyruvate metabolism pathway in the liver. These results indicate the promising prospect of using LcS for the treatment of liver diseases, particularly ALI.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"15 1","pages":"247-261"},"PeriodicalIF":5.7,"publicationDate":"2021-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13750","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5711459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microbial production of small peptide: pathway engineering and synthetic biology","authors":"Zhiyong Wu,&nbsp;Youran Li,&nbsp;Liang Zhang,&nbsp;Zhongyang Ding,&nbsp;Guiyang Shi","doi":"10.1111/1751-7915.13743","DOIUrl":"https://doi.org/10.1111/1751-7915.13743","url":null,"abstract":"<p>Small peptides are a group of natural products with low molecular weights and complex structures. The diverse structures of small peptides endow them with broad bioactivities and suggest their potential therapeutic use in the medical field. The remaining challenge is methods to address the main limitations, namely (i) the low amount of available small peptides from natural sources, and (ii) complex processes required for traditional chemical synthesis. Therefore, harnessing microbial cells as workhorse appears to be a promising approach to synthesize these bioactive peptides. As an emerging engineering technology, synthetic biology aims to create standard, well-characterized and controllable synthetic systems for the biosynthesis of natural products. In this review, we describe the recent developments in the microbial production of small peptides. More importantly, synthetic biology approaches are considered for the production of small peptides, with an emphasis on chassis cells, the evolution of biosynthetic pathways, strain improvements and fermentation.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 6","pages":"2257-2278"},"PeriodicalIF":5.7,"publicationDate":"2021-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13743","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5699676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aspergillus fumigatus AR04 obeys Arrhenius' rule in cultivation temperature shifts from 30 to 40°C","authors":"Susanne Nieland,&nbsp;Susann Barig,&nbsp;Julian Salzmann,&nbsp;Frauke Gehrau,&nbsp;Arief Izzairy Zamani,&nbsp;Annabell Richter,&nbsp;Julia Ibrahim,&nbsp;Yvonne Gr?ser,&nbsp;Chyan Leong Ng,&nbsp;Klaus-Peter Stahmann","doi":"10.1111/1751-7915.13739","DOIUrl":"https://doi.org/10.1111/1751-7915.13739","url":null,"abstract":"<p>To set a benchmark in fungal growth rate, a differential analysis of prototrophic <i>Aspergillus fumigatus</i> AR04 with three ascomycetes applied in &gt; 10³ t year^-1 scale was performed, i.e. <i>Ashbya gosspyii</i> (riboflavin), <i>Aspergillus niger</i> (citric acid) and <i>Aspergillus oryzae</i> (food-processing). While radial colony growth decreased 0.5-fold when <i>A</i>. <i>gossypii</i> was cultivated at 40°C instead of 28°C, <i>A. fumigatus</i> AR04 responded with 1.7-fold faster hyphal growth. <i>A</i>. <i>niger</i> and <i>A. oryzae</i> formed colonies at 40°C, but not at 43°C. Moreover, all <i>A. fumigatus</i> strains tested grew even at 49°C. In chemostat experiments, <i>A</i>. <i>fumigatus</i> AR04 reached steady state at a dilution rate of 0.7 h^-1 at 40°C, 120% more than reported for <i>A. gossypii</i> at 28°C. To study mycelial growth rates under unlimited conditions, carbon dioxide increase rates were calculated from concentrations detected online in the exhaust of batch fermentations for 3 h only. All rates calculated suggest that <i>A. fumigatus</i> AR04 approximates Arrhenius’ rule when comparing short cultivations at 30°C with those at 40°C. Linearization of the exponential phase and comparison of the slopes revealed an increase to 192% by the 10°C up-shift.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 4","pages":"1422-1432"},"PeriodicalIF":5.7,"publicationDate":"2021-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13739","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6164591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome analysis of the metabolically versatile Pseudomonas umsongensis GO16: the genetic basis for PET monomer upcycling into polyhydroxyalkanoates","authors":"Tanja Narancic,&nbsp;Manuel Salvador,&nbsp;Graham M. Hughes,&nbsp;Niall Beagan,&nbsp;Umar Abdulmutalib,&nbsp;Shane T. Kenny,&nbsp;Huihai Wu,&nbsp;Marta Saccomanno,&nbsp;Jounghyun Um,&nbsp;Kevin E. O'Connor,&nbsp;José I. Jiménez","doi":"10.1111/1751-7915.13712","DOIUrl":"https://doi.org/10.1111/1751-7915.13712","url":null,"abstract":"<p>The throwaway culture related to the single-use materials such as polyethylene terephthalate (PET) has created a major environmental concern. Recycling of PET waste into biodegradable plastic polyhydroxyalkanoate (PHA) creates an opportunity to improve resource efficiency and contribute to a circular economy. We sequenced the genome of <i>Pseudomonas umsongensis</i> GO16 previously shown to convert PET-derived terephthalic acid (TA) into PHA and performed an in-depth genome analysis. GO16 can degrade a range of aromatic substrates in addition to TA, due to the presence of a catabolic plasmid pENK22. The genetic complement required for the degradation of TA <i>via</i> protocatechuate was identified and its functionality was confirmed by transferring the <i>tph</i> operon into <i>Pseudomonas putida</i> KT2440, which is unable to utilize TA naturally. We also identified the genes involved in ethylene glycol (EG) metabolism, the second PET monomer, and validated the capacity of GO16 to use EG as a sole source of carbon and energy. Moreover, GO16 possesses genes for the synthesis of both medium and short chain length PHA and we have demonstrated the capacity of the strain to convert mixed TA and EG into PHA. The metabolic versatility of GO16 highlights the potential of this organism for biotransformations using PET waste as a feedstock.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 6","pages":"2463-2480"},"PeriodicalIF":5.7,"publicationDate":"2021-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13712","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6115385","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LoxTnSeq: random transposon insertions combined with cre/lox recombination and counterselection to generate large random genome reductions","authors":"Daniel Shaw,&nbsp;Samuel Miravet-Verde,&nbsp;Carlos Pi?ero-Lambea,&nbsp;Luis Serrano,&nbsp;Maria Lluch-Senar","doi":"10.1111/1751-7915.13714","DOIUrl":"https://doi.org/10.1111/1751-7915.13714","url":null,"abstract":"<p>The removal of unwanted genetic material is a key aspect in many synthetic biology efforts and often requires preliminary knowledge of which genomic regions are dispensable. Typically, these efforts are guided by transposon mutagenesis studies, coupled to deepsequencing (TnSeq) to identify insertion points and gene essentiality. However, epistatic interactions can cause unforeseen changes in essentiality after the deletion of a gene, leading to the redundancy of these essentiality maps. Here, we present LoxTnSeq, a new methodology to generate and catalogue libraries of genome reduction mutants. LoxTnSeq combines random integration of lox sites by transposon mutagenesis, and the generation of mutants via Cre recombinase, catalogued via deep sequencing. When LoxTnSeq was applied to the naturally genome reduced bacterium <i>Mycoplasma pneumoniae</i>, we obtained a mutant pool containing 285 unique deletions. These deletions spanned from &gt; 50 bp to 28 Kb, which represents 21% of the total genome. LoxTnSeq also highlighted large regions of non-essential genes that could be removed simultaneously, and other non-essential regions that could not, providing a guide for future genome reductions.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 6","pages":"2403-2419"},"PeriodicalIF":5.7,"publicationDate":"2020-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13714","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5682100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The sulfur formation system mediating extracellular cysteine-cystine recycling in Fervidobacterium islandicum AW-1 is associated with keratin degradation","authors":"Hyeon-Su Jin,&nbsp;Immanuel Dhanasingh,&nbsp;Jae-Yoon Sung,&nbsp;Jae Won La,&nbsp;Yena Lee,&nbsp;Eun Mi Lee,&nbsp;Yujin Kang,&nbsp;Do Yup Lee,&nbsp;Sung Haeng Lee,&nbsp;Dong-Woo Lee","doi":"10.1111/1751-7915.13717","DOIUrl":"https://doi.org/10.1111/1751-7915.13717","url":null,"abstract":"<p>Most extremophilic anaerobes possess a sulfur formation (Suf) system for Fe–S cluster biogenesis. In addition to its essential role in redox chemistry and stress responses of Fe–S cluster proteins, the Suf system may play an important role in keratin degradation by <i>Fervidobacterium islandicum</i> AW-1. Comparative genomics of the order Thermotogales revealed that the feather-degrading <i>F. islandicum</i> AW-1 has a complete Suf-like machinery (SufCBDSU) that is highly expressed in cells grown on native feathers in the absence of elemental sulfur (S⁰). On the other hand, <i>F. islandicum</i> AW-1 exhibited a significant retardation in the Suf system-mediated keratin degradation in the presence of S⁰. Detailed differential expression analysis of sulfur assimilation machineries unveiled the mechanism by which an efficient sulfur delivery from persulfurated SufS to SufU is achieved during keratinolysis under sulfur starvation. Indeed, addition of SufS–SufU to cell extracts containing keratinolytic proteases accelerated keratin decomposition <i>in vitro</i> under reducing conditions. Remarkably, mass spectrometric analysis of extracellular and intracellular levels of amino acids suggested that redox homeostasis within cells coupled to extracellular cysteine and cystine recycling might be a prerequisite for keratinolysis. Taken together, these results suggest that the Suf-like machinery including the SufS–SufU complex may contribute to sulfur availability for an extracellular reducing environment as well as intracellular redox homeostasis through cysteine released from keratin hydrolysate under starvation conditions.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 3","pages":"938-952"},"PeriodicalIF":5.7,"publicationDate":"2020-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13717","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6350910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reconstitution of a mini-gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus","authors":"Dong Li,&nbsp;Yuqing Tian,&nbsp;Xiang Liu,&nbsp;Wenxi Wang,&nbsp;Yue Li,&nbsp;Huarong Tan,&nbsp;Jihui Zhang","doi":"10.1111/1751-7915.13686","DOIUrl":"https://doi.org/10.1111/1751-7915.13686","url":null,"abstract":"<p>Salinomycin, an FDA-approved polyketide drug, was recently identified as a promising anti-tumour and anti-viral lead compound. It is produced by <i>Streptomyces albus</i>, and the biosynthetic gene cluster (<i>sal</i>) spans over 100 kb. The genetic manipulation of large polyketide gene clusters is challenging, and approaches delivering reliable efficiency and accuracy are desired. Herein, a delicate strategy to enhance salinomycin production was devised and evaluated. We reconstructed a minimized <i>sal</i> gene cluster (mini-cluster) on pSET152 including key genes responsible for tailoring modification, antibiotic resistance, positive regulation and precursor supply. These genes were overexpressed under the control of constitutive promoter P<i>_kasO*</i> or P<i>_neo</i>. The <i>pks</i> operon was not included in the mini-cluster, but it was upregulated by SalJ activation. After the plasmid pSET152::mini-cluster was introduced into the wild-type strain and a chassis host strain obtained by ribosome engineering, salinomycin production was increased to 2.3-fold and 5.1-fold compared with that of the wild-type strain respectively. Intriguingly, mini-cluster introduction resulted in much higher production than overexpression of the whole <i>sal</i> gene cluster. The findings demonstrated that reconstitution of <i>sal</i> mini-cluster combined with ribosome engineering is an efficient novel approach and may be extended to other large polyketide biosynthesis.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 6","pages":"2356-2368"},"PeriodicalIF":5.7,"publicationDate":"2020-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13686","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6071189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Channelling carbon flux through the meta-cleavage route for improved poly(3-hydroxyalkanoate) production from benzoate and lignin-based aromatics in Pseudomonas putida H","authors":"José Manuel Borrero-de Acu?a,&nbsp;Izabook Gutierrez-Urrutia,&nbsp;Cristian Hidalgo-Dumont,&nbsp;Carla Aravena-Carrasco,&nbsp;Matias Orellana-Saez,&nbsp;Nestor Palominos-Gonzalez,&nbsp;Jozef B. J. H. van Duuren,&nbsp;Viktoria Wagner,&nbsp;Lars Gl?ser,&nbsp;Judith Becker,&nbsp;Michael Kohlstedt,&nbsp;Flavia C. Zacconi,&nbsp;Christoph Wittmann,&nbsp;Ignacio Poblete-Castro","doi":"10.1111/1751-7915.13705","DOIUrl":"https://doi.org/10.1111/1751-7915.13705","url":null,"abstract":"<p>Lignin-based aromatics are attractive raw materials to derive medium-chain length poly(3-hydroxyalkanoates) (<i>mcl</i>-PHAs), biodegradable polymers of commercial value. So far, this conversion has exclusively used the <i>ortho-</i>cleavage route of <i>Pseudomonas putida</i> KT2440, which results in the secretion of toxic intermediates and limited performance. <i>Pseudomonas putida</i> H exhibits the <i>ortho-</i> and the <i>meta-</i>cleavage pathways where the latter appears promising because it stoichiometrically yields higher levels of acetyl-CoA. Here, we created a double-mutant H-Δ<i>catA</i>Δ<i>A2</i> that utilizes the <i>meta</i> route exclusively and synthesized 30% more PHA on benzoate than the parental strain but suffered from catechol accumulation. The single deletion of the <i>catA</i>2 gene in the H strain provoked a slight attenuation on the enzymatic capacity of the <i>ortho</i> route (25%) and activation of the <i>meta</i> route by nearly 8-fold, producing twice as much <i>mcl</i>-PHAs compared to the wild type. Inline, the mutant H-Δ<i>catA</i>2 showed a 2-fold increase in the intracellular malonyl-CoA abundance – the main precursor for <i>mcl-</i>PHAs synthesis. As inferred from flux simulation and enzyme activity assays, the superior performance of H-Δ<i>catA</i>2 benefited from reduced flux through the TCA cycle and malic enzyme and diminished by-product formation. In a benzoate-based fed-batch, <i>P</i>. <i>putida</i> H-Δ<i>catA</i>2 achieved a PHA titre of 6.1 g l^–1 and a volumetric productivity of 1.8 g l^–1 day^–1. Using Kraft lignin hydrolysate as feedstock, the engineered strain formed 1.4 g l^- 1 PHA. The balancing of carbon flux between the parallel catechol-degrading routes emerges as an important strategy to prevent intermediate accumulation and elevate <i>mcl-</i>PHA production in <i>P. putida</i> H and, as shown here, sets the next level to derive this sustainable biopolymer from lignin hydrolysates and aromatics.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 6","pages":"2385-2402"},"PeriodicalIF":5.7,"publicationDate":"2020-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13705","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6192404","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deletion of a hybrid NRPS-T1PKS biosynthetic gene cluster via Latour gene knockout system in Saccharopolyspora pogona and its effect on butenyl-spinosyn biosynthesis and growth development","authors":"Jie Rang,&nbsp;Yunlong Li,&nbsp;Li Cao,&nbsp;Ling Shuai,&nbsp;Yang Liu,&nbsp;Haocheng He,&nbsp;Qianqian Wan,&nbsp;Yuewen Luo,&nbsp;Ziquan Yu,&nbsp;Youming Zhang,&nbsp;Yunjun Sun,&nbsp;Xuezhi Ding,&nbsp;Shengbiao Hu,&nbsp;Qingji Xie,&nbsp;Liqiu Xia","doi":"10.1111/1751-7915.13694","DOIUrl":"https://doi.org/10.1111/1751-7915.13694","url":null,"abstract":"<p>Butenyl-spinosyn, a promising biopesticide produced by <i>Saccharopolyspora pogona</i>, exhibits stronger insecticidal activity and a broader pesticidal spectrum. However, its titre in the wild-type <i>S. pogona</i> strain is too low to meet the industrial production requirements. Deletion of non-target natural product biosynthetic gene clusters resident in the genome of <i>S. pogona</i> could reduce the consumption of synthetic precursors, thereby promoting the biosynthesis of butenyl-spinosyn. However, it has always been a challenge for scientists to genetically engineer <i>S. pogona</i>. In this study, the Latour gene knockout system (linear DNA fragment recombineering system) was established in <i>S. pogona</i>. Using the Latour system, a hybrid NRPS-T1PKS cluster (˜20 kb) which was responsible for phthoxazolin biosynthesis was efficiently deleted in <i>S. pogona</i>. The resultant mutant <i>S. pogona</i>-Δ<i>ura4-</i>Δc14 exhibited an extended logarithmic phase, increased biomass and a lower glucose consumption rate. Importantly, the production of butenyl-spinosyn in <i>S. pogona</i>-Δ<i>ura4-</i>Δc14 was increased by 4.72-fold compared with that in the wild-type strain. qRT-PCR analysis revealed that phthoxazolin biosynthetic gene cluster deletion could promote the expression of the butenyl-spinosyn biosynthetic gene cluster. Furthermore, a TetR family transcriptional regulatory gene that could regulate the butenyl-spinosyn biosynthesis has been identified from the phthoxazolin biosynthetic gene cluster. Because dozens of natural product biosynthetic gene clusters exist in the genome of <i>S. pogona</i>, the strategy reported here will be used to further promote the production of butenyl-spinosyn by deleting other secondary metabolite synthetic gene clusters.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"14 6","pages":"2369-2384"},"PeriodicalIF":5.7,"publicationDate":"2020-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13694","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5910732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Screening of a genome-reduced Corynebacterium glutamicum strain library for improved heterologous cutinase secretion","authors":"Johannes Hemmerich,&nbsp;Mohamed Labib,&nbsp;Carmen Steffens,&nbsp;Sebastian J. Reich,&nbsp;Marc Weiske,&nbsp;Meike Baumgart,&nbsp;Christian Rückert,&nbsp;Matthias Ruwe,&nbsp;Daniel Siebert,&nbsp;Volker F. Wendisch,&nbsp;J?rn Kalinowski,&nbsp;Wolfgang Wiechert,&nbsp;Marco Oldiges","doi":"10.1111/1751-7915.13660","DOIUrl":"https://doi.org/10.1111/1751-7915.13660","url":null,"abstract":"<p>The construction of microbial platform organisms by means of genome reduction is an ongoing topic in biotechnology. In this study, we investigated whether the deletion of single or multiple gene clusters has a positive effect on the secretion of cutinase from <i>Fusarium solani pisi</i> in the industrial workhorse <i>Corynebacterium glutamicum</i>. A total of 22 genome-reduced strain variants were compared applying two Sec signal peptides from <i>Bacillus subtilis</i>. High-throughput phenotyping using robotics-integrated microbioreactor technology with automated harvesting revealed distinct cutinase secretion performance for a specific combination of signal peptide and genomic deletions. The biomass-specific cutinase yield for strain GRS41_51_NprE was increased by ~ 200%, although the growth rate was reduced by ~ 60%. Importantly, the causative deletions of genomic clusters cg2801-cg2828 and <i>rrnC</i>-cg3298 could not have been inferred <i>a priori</i>. Strikingly, bioreactor fed-batch cultivations at controlled growth rates resulted in a complete reversal of the screening results, with the cutinase yield for strain GRS41_51_NprE dropping by ~ 25% compared to the reference strain. Thus, the choice of bioprocess conditions may turn a ‘high-performance’ strain from batch screening into a ‘low-performance’ strain in fed-batch cultivation. In conclusion, future studies are needed in order to understand metabolic adaptations of <i>C. glutamicum</i> to both genomic deletions and different bioprocess conditions.</p>","PeriodicalId":49145,"journal":{"name":"Microbial Biotechnology","volume":"13 6","pages":"2020-2031"},"PeriodicalIF":5.7,"publicationDate":"2020-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/1751-7915.13660","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5737853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}