Journal of Industrial Microbiology & Biotechnology最新文献

{"title":"Complete diploid genome of the type strain Yarrowia lipolytica YB-423 (ATCC 18942™).","authors":"James E Crill, Scott V Nguyen, Corina Tabron, Nikhita Puthuveetil, Anthony Muhle, Zethus W Avery, Joseph Petrone, Kaitlyn Bentley, Jade Kirkland, Noah Wax, Robert Marlow, James Duncan, Steve King, Ana Fernandes, John Bagnoli, Briana Benton, Shahin S Ali, Roy D Welch, Jonathan L Jacobs","doi":"10.1093/jimb/kuag002","DOIUrl":"10.1093/jimb/kuag002","url":null,"abstract":"<p><p>Here, we present the first complete, fully phased diploid genome of type strain Yarrowia lipolytica YB-423 (=ATCC 18942™), constructed using a combination of Oxford Nanopore long-read and Illumina short-read sequencing. Yarrowia lipolytica is an industrially relevant yeast species known for its metabolic versatility, particularly its ability to degrade hydrophobic compounds and express useful products such as fatty acids. Despite its growing use in biotechnology, a high-quality genome assembly of the species' diploid type-strain has been lacking. The assembly and annotations presented here span six chromosomes of paired \"haplotigs\" and a mitochondrial genome, capturing large-scale structural variations and prominent levels of genome-wide heterozygosity. Variant analysis revealed 13,908 heterozygous alleles, of which 3,201 alleles were distributed among 1,237 protein-coding genes. Gene set enrichment analysis showed that these variants are enriched among genes involved in transmembrane transport, suggesting a role in environmental adaptability. Comparative analysis of matched haplotigs for the same chromosome uncovered multiple inversions and transpositions, as well as allele-specific insertions of retrotransposons, providing new insights into the structural complexity and evolutionary dynamics of the genome. The fully phased, finished diploid genome of Y. lipolytica YB-423 represents a crucial step toward unlocking the full genetic potential of Y. lipolytica. Our work will provide a valuable foundation for future comparative and functional genomics and strain engineering studies, particularly for industrial microbiology and biotechnology applications. One-sentence summary This study presents the first fully phased diploid genome of Yarrowia lipolytica type strain YB-423, revealing extensive structural variation and heterozygosity that enhance understanding of its genetic adaptability and industrial potential.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12888388/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145933622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Statistical approach for media optimization of fHbp-PorA chimeric protein expression in Escherichia coli; development of a promising Meningococcal B vaccine.","authors":"Annamraju Aswini, Annamraju D Sarma, Sambhaji S Pisal, Selvan Ravindran","doi":"10.1093/jimb/kuag001","DOIUrl":"10.1093/jimb/kuag001","url":null,"abstract":"<p><p>Successful design of an industrial biological product such as a vaccine requires an efficient, robust and reproducible process. In this study, we investigated conditions for process optimization of a promising recombinant factor H binding protein (fHbp)-Porin A (PorA) chimeric protein-based vaccine candidate against Meningococcal B serogroup in Escherichia coli B834 strain using the inducible T7-lac promoter system. Random screening of components of complex culture media for growth and expression using IPTG as an inducer, showed inconsistency when analyzed using Plackett-Burman design (PBD). Further analysis identified galactose present either in tryptone (as lactose) or soy-phytone, has caused strong autoinduction and is surmised as an interfering factor for IPTG induction. Synergistic combination of soy-phytone and yeast extract under conditions of controlled growth and auto-induction gave a good correlation between the cell growth and expression of the fHbp-PorA chimeric protein when evaluated using PBD and central composite design. Using response optimization conditions, an optimized media was attained. A shake flask study was performed to validate the optimized media, and later, a fed-batch fermentation at 10 L scale was established to prove the scalability, consistency, and product quality using the optimized media. One-sentence summary This study reports optimization of cell growth and expression conditions for a recombinant chimeric meningococcal protein in E. coli that assures its industrial scale production and suitability for preclinical and clinical studies as a vaccine component against Meningococcal B serogroup.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12883990/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145933617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synergistic strategies for high production of Geobacillus stearothermophilus α-amylase in Bacillus subtilis.","authors":"Deming Rao, Changhui Wang, Xiaolin Li, Wei Shen, Qiang Liu, Zerui Li, Shiyu Pi, Zhenggang Han, Jiangke Yang","doi":"10.1093/jimb/kuaf036","DOIUrl":"10.1093/jimb/kuaf036","url":null,"abstract":"<p><p>Although the α-amylase (AmyS) from Geobacillus stearothermophilus exhibits high thermostability, its low enzymatic activity (44.36 ± 2.02 U/mL) severely hinders industrial applications. Given its strong protein secretion capability and GRAS (generally recognized as safe) status, this study employed Bacillus subtilis as the host to enhance AmyS production. Through error-prone PCR and high-throughput screening, a triple mutant T151A/K178E/T458A (AmySM) was generated, showing a 17.67% increase in activity (51.34 ± 1.11 U/mL). AmySM activity was further increased by 29.32% to 65.23 ± 2.33 U/mL using the signal peptide SPykwD, selected from a comprehensive library for superior secretion efficiency. The promoter PgsiB enhanced activity by 37.61% to 89.54 ± 2.95 U/mL. Optimizing the ribosome binding site (RBS) resulted in an additional 48.83% increase in activity, yielding a final activity of 134.02 ± 3.54 U/mL, which corresponds to a 3.02-fold improvement over the initial strain WBSW (44.36 ± 2.02 U/mL). Ultimately, scale-up fermentation in a 5-L bioreactor yielded a maximum extracellular activity of 1244.17 ± 48.66 U/mL at 72 hours, a 2.84-fold increase over the control. This multi-level strategy provides a rational framework for high-efficiency AmySM production, paving the way for extracellular production of high-value proteins in the GRAS host B. subtilis.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12784944/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145794179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Laboratory and field performance of 2-(thiocyanomethylthio)benzothiazole against fast-growing nontuberculous mycobacteria found in metalworking fluid.","authors":"Christopher L Wiatr, Carey Cribbs, C B French","doi":"10.1093/jimb/kuag019","DOIUrl":"10.1093/jimb/kuag019","url":null,"abstract":"<p><p>Uncontrolled, fast-growing nontuberculous mycobacteria (NTM) in metalworking fluids (MWFs) can cause dermal lesions, hypersensitivity pneumonitis, and other health complications in exposed metalworkers. Although biocides are routinely used to control microbial growth in MWF, they are often ineffective because they are unsuitable for the physical or chemical conditions of the system, added infrequently, or ineffective against the microorganisms present. Given the increasing concern for NTM in industrial settings, there is a growing need for MWF-compatible (i.e., nonoxidizing) biocides that are effective against NTM. Here, we evaluated the efficacy of four nonoxidizing biocides against fast-growing NTM species in water-based MWF. Results from a 72-hr laboratory challenge indicated that biocides containing 2-(thiocyanomethylthio)benzothiazole (TCMTB), commonly used as a fungicide, were most effective at controlling Mycobacterium chelonae, M. fortuitum, and field and laboratory strains of M. immunogenum. In subsequent field trials at nine MWF plants where workers had reported symptoms of dermal and lung illnesses, TCMTB consistently reduced viable NTM counts, as measured through both culture-based assays and microscopy. However, dosages needed to be increased two- to four-fold relative to the laboratory experiments to control NTM populations in systems with established biofilms. Due to the observed success of TCMTB against NTM in field context, the manufacturing plants involved in this study continued regular application of TCMTB as a preventative treatment. No additional outbreaks were observed throughout our follow-up period, which, in several cases, spanned approximately 10 years. The results suggest that TCMTB may provide an efficacious method for treating and preventing NTM contamination in MWF systems. Future studies should explore the mechanism of action of the sulfur-rich chemical structure of TCMTB to potentially guide the development and synthesis of related compounds with anti-NTM properties. One sentence summary 2--(thiocyanomethylthio)benzothiazole kills fast-growing nontuberculous mycobacteria, including Mycobacterium immunogenum, and prevents worker infections based on laboratory, field trial, and long-term (6-10 years) industrial studies.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13446119/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148471081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation and characterization of Saccharomyces cerevisiae mutants with ornithine accumulation for value-added craft beer brewing.","authors":"Akira Nishimura, Shota Isogai, Koya Yamada, Ryoya Tanahashi, Hiroshi Takagi","doi":"10.1093/jimb/kuag013","DOIUrl":"10.1093/jimb/kuag013","url":null,"abstract":"<p><p>Ornithine is a nonproteinogenic amino acid with various health benefits, making it a promising target for functional food development. In this study, we developed the yeast Saccharomyces cerevisiae strains with elevated intracellular ornithine and identified the genetic mutation responsible for this trait. An ornithine-rich mutant was successfully isolated by chemical mutagenesis and selection for resistance to canavanine, a toxic arginine analog. Whole-genome sequencing revealed a heterozygous point mutation in the ARG6 gene encoding N-acetylglutamate kinase, a key enzyme in ornithine biosynthesis. This mutation caused a Gly351Asp substitution located in a conserved linker region between functional domains. Introduction of this substitution, along with introducing alternative amino acids at the same site, consistently increased ornithine levels in various strain backgrounds. Structural modeling suggested that this substitution could affect the enzyme conformation or inter-domain interactions. These results establish a practical nongenetically modified breeding strategy for enhancing ornithine production in yeast, which will facilitate the development of ornithine-enriched fermented beverages such as craft beer. One-sentence summary The ARG6 mutant increases intracellular ornithine and enables value-added craft beer brewing.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13261765/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147973014","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating the efficacy of disinfecting products against Bacillus subtilis spore using an optimized method.","authors":"Yoonsung Hu, Jinshan Jin, Haijing Hu, Ian F Deveau, Steven L Foley, Huizhong Chen","doi":"10.1093/jimb/kuag025","DOIUrl":"10.1093/jimb/kuag025","url":null,"abstract":"<p><p>Twenty disinfectant products were evaluated for sporicidal activity using an optimized method. Bacillus subtilis ATCC 6051 was sporulated in Difco sporulation medium for 3 days. The spores were purified using density centrifugation and heat treatment and then prepared for sporicidal efficacy assessment. The quality of the purified spores was verified using sodium hypochlorite solutions with free chlorine concentrations of 1,500 and 5,000 ppm. The active ingredients in disinfectant products included chlorine, hydrogen peroxide/peracetic acid, aldehydes, quaternary ammonium compounds, and phenols. Of the 20 products tested, 6 met the effectiveness criterion of greater than a 5 log₁₀ reduction after exposure of B. subtilis spores for the label-recommended contact time. Among the 11 products claiming sporicidal activity, 4 met the established criterion. Among the five categories of active ingredients evaluated, some chlorine-based and hydrogen peroxide/peracetic acid-based products demonstrated significantly higher sporicidal activity than products in the other three categories. Overall, products that did not meet the study criteria exhibited less than a 1 log₁₀ reduction. When these underperforming sporicidal products were tested with an extended contact time of 10 min, one additional product met the effectiveness criterion. Subsequent quality assessments using freshly prepared, laboratory-made products containing identical active ingredients demonstrated improved sporicidal efficacy compared with the original test items, indicating that defects or degradation of active ingredients may have occurred in some of the tested commercial disinfectant products. One sentence summary Evaluation of 20 disinfectants against Bacillus subtilis spores using optimized methods.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13548849/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148794147","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expanding the genetic toolset: using serine recombinases to integrate riboregulatory elements into industrially relevant microbial chassis.","authors":"Katherine L Wozniak, Austin L Carroll, Steffi A Davison, Ramesh K Jha, Adam M Guss, Scott P Hennelly, Taraka Dale, Chris M Yeager","doi":"10.1093/jimb/kuag015","DOIUrl":"10.1093/jimb/kuag015","url":null,"abstract":"<p><p>To realize the full potential of biomanufacturing, the breadth of industrial microbes used to consume diverse feedstock and generate bioproducts needs to expand. As such, portable tools are required that can be used by multiple hosts for straightforward genomic manipulation and precise gene expression. Here, we demonstrate the co-utilization of two synthetic biology tools to achieve these goals: cis-repressors (CRs) and serine recombinase-assisted genome engineering (SAGE). CRs are small, noncoding RNAs that are placed upstream of the target gene to modulate bacterial translation rates at varying, discrete levels. SAGE uses site-specific serine recombinases to catalyze highly efficient, unidirectional insertion of DNA into the chromosome of diverse organisms. We used SAGE to integrate a suite of CRs into the industrially relevant hosts Pseudomonas putida, Corynebacterium glutamicum, and Cupriavidus necator. Using a fluorescent reporter as a readout of CR functionality, we found that CR performance across these backgrounds was similar-providing a range of translational repression up to 100-fold. Overall, these results demonstrate the high portability of CRs across bacterial genetic backgrounds, which ideally can be used in future microbial engineering efforts pertinent to biomanufacturing. One-Sentence Summary: Fine tuning translation across phylogenetically diverse microorganisms using riboregulators and serine recombinase-assisted genome engineering.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13289732/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148149718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Innovations in process engineering approaches for enhanced bacteriocin production.","authors":"Joan Sweetlin Shanmugam, Sreeja Shanmuga Doss","doi":"10.1093/jimb/kuag010","DOIUrl":"10.1093/jimb/kuag010","url":null,"abstract":"<p><p>Bacteriocins are ribosomally synthesized antimicrobial peptides produced by various classes of bacteria, exhibiting broad-spectrum activity that makes them promising candidates for applications in food preservation and medicine. Their inherent stability under extreme pH, temperature, and salinity conditions further supports their functional versatility. However, the widespread industrial application of bacteriocins remains constrained by the low titres typically achieved during fermentation. Despite extensive efforts to optimize production using batch and fed-batch fermentation strategies, the resulting titres remain inadequate for economically viable large-scale manufacturing. This review aims to provide a comprehensive overview of novel process strategies developed over the past two decades to enhance bacteriocin yields during fermentation. One of the primary challenges is the inhibition of microbial growth due to the accumulation of lactic acid or the bacteriocin itself during production. To address this, in situ product removal techniques-such as co-cultivation with lactic acid-consuming microorganisms, in situ adsorption, filtration, and foam fractionation-have been explored, yielding notable improvements in bacteriocin titres. Additionally, stress-induced production strategies involving biological (e.g., co-culture with competing microbes), chemical (e.g., salinity and pH stress), and physical (e.g., agitation, temperature, and aeration stress) stimuli have also demonstrated success in enhancing bacteriocin synthesis. This review underscores the importance of these innovative fermentation approaches and highlights the need for further research focused on scaling up such processes. Advancing these strategies is critical to realizing the full potential of bacteriocins in food safety, antimicrobial therapy, and broader biomedical applications. One-sentence summary This review provides a comprehensive view of the novel fermentation strategies developed in the last two decades to overcome low bacteriocin titres during fermentation such as in situ lactate removal, in situ bacteriocin removal, and stress-led induction.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13037702/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147468193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Butyric acid production from corn: economics of fermentative production of a novel biobased chemical.","authors":"Nasib Qureshi, Somesh Mishra, Zengshe Liu, Mukti Singh, Ronald E Hector, Thaddeus C Ezeji, Richard D Ashby, Michael A Jackson, Vijay Singh","doi":"10.1093/jimb/kuag009","DOIUrl":"10.1093/jimb/kuag009","url":null,"abstract":"<p><p>In the present study, the economic evaluation of fermentation produced butyric acid (HBu) from corn was carried out (capacity 40 × 103 metric tons HBu∙yr-1) and two recovery processes (distillation and adsorption) were compared. Distillation is a longstanding process that has been effectively used to recover volatile chemicals. Recently, we developed an adsorption process that can be used to recover HBu cost competitively. The direct fixed capital (DFC) of the two processes (distillation & adsorption) were (in US dollars) $73.96 × 106 and $43.91 × 106, respectively. It is reported that the production of HBu from corn requires supplementing with amylase enzymes to the corn mash. For the two processes (distillation & adsorption) the annual operating costs were $37.67 × 106 and $31.83 × 106, respectively. The utilities annual costs for the two processes were $47.38 × 106 and $5.91 × 106, respectively. Corn price is one of the most important factors that influence HBu selling price, but other factors that could lower production costs include plant capacity. Factors such as tax on profit, interest rates on borrowed capital and plant life have marginal or low impact on HBu selling price. The significance of this paper is that HBu produced from corn and recovered by adsorption could be sold for $1.22 to 0.85∙kg-1. The selling price of fermentation produced HBu could further be reduced considerably if agricultural biomass such as wheat straw is used for production. One sentence summary Butyric acid production by fermentation and process economics.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13070389/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147443780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multistrategy metabolic engineering of Talaromyces pinophilus for α-amylase production from lignocellulosic biomass.","authors":"Jing Zeng, Jianjun Guo, Shuaiwen Zhang, Ya Li, Siyuan Yue, Peng Li, Junhui Nie, Tong Wang, Cheng Zhang, Lin Yuan","doi":"10.1093/jimb/kuag021","DOIUrl":"10.1093/jimb/kuag021","url":null,"abstract":"<p><p>Filamentous fungi are important hosts for industrial enzyme production. Growing demand for α-amylase has increased reliance on food-derived carbon substrates, necessitating fungal strains that efficiently utilize nongrain biomass. In this study, Talaromyces pinophilus Y117 was metabolically engineered to produce α-amylase from lignocellulosic biomass. A strong cellobiohydrolase I gene (cbh1) promoter (Pcbh1Tru) was identified to drive expression. Multiple rounds of multilocus integration of the α-amylase gene were performed using homologous multicopy genomic sequences as recombination arms with a Cre/loxP-based recyclable selection system, yielding the multicopy strain Tp4, which achieved 4124.5 U/mL α-amylase activity in shake-flask fermentation with corncob powder as the sole carbon source. To minimize enzyme degradation, the protease gene 8538 was deleted using the Cre/lox2272 system, generating Tp4Δp. This strain showed a 50% increase in shake-flask α-amylase activity (6208.4 U/mL). In 3-L bioreactor cultivation, Tp4Δp exhibited excellent production performance, achieving 26 712.2 U/mL α-amylase activity. When corncob powder was used as the sole substrate, the cellulose and hemicellulose degradation rates reached 90.00% and 70.01%, respectively, and the enzyme yield reached 213 697.5 U per gram of corncob powder. This engineered strain demonstrates strong potential for industrial applications. The synthesis-degradation synergistic optimization strategy provides a practical approach for engineering filamentous fungal cell factories to produce enzymes directly from lignocellulosic biomass. One sentence summary Metabolic engineering of Talaromyces pinophilus through promoter optimization, multicopy integration, and protease deletion enables efficient α-amylase production from lignocellulosic biomass, achieving 26 712 U/mL in bioreactor fermentation.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13487897/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148794135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}