Applied Microbiology and Biotechnology最新文献

{"title":"A neutralizing nanobody-based liquid-phase blocking ELISA to assess the protective potency of Senecavirus A vaccine","authors":"Suyu Mu,&nbsp;Songjia Pan,&nbsp;Hu Dong,&nbsp;Jinen Wu,&nbsp;Yun Zhang,&nbsp;Shuanghui Yin,&nbsp;Jianing Wang,&nbsp;Tian Wei,&nbsp;Xiaobo Wen,&nbsp;Huichen Guo,&nbsp;Shiqi Sun","doi":"10.1007/s00253-025-13492-4","DOIUrl":"10.1007/s00253-025-13492-4","url":null,"abstract":"<p><i>Senecavirus</i> A (SVA) causes swine blister diseases in the crown of hooves, abortion syndrome, and increased mortality rates in piglets. Vaccination is the best defense against SVA infection. However, evaluations of the efficacy of SVA vaccines currently rely on challenging pigs with SVA, which is time-consuming, labor-intensive, and inconsistent with animal welfare mandates. To establish a replacement for the virus neutralization test (VNT) and SVA challenge tests, we developed a liquid-phase blocking enzyme-linked immunosorbent assay (nbLPB-ELISA) based on V1-VHH as the coating antibody (Ab) and biotinylated V1-VHH as the detection Ab. Under optimized conditions, the VNT and nbLPB-ELISA results were strongly correlated (Pearson <i>R</i>² = 0.84; <i>p</i> &lt; 0.00001). Analysis of the LBP-ELISA and vaccine protection rate revealed that neutralizing Ab titers greater than 256 provided 100% protection, while titers of 64 and 128 offered 57.1% and 70% protection, respectively. The nbLPB-ELISA is a rapid, simple, safe, and cost-effective method of detecting SVA as a replacement for the SVA VNT and vaccine-challenge assays.</p><p><i>• We evaluated a specific, high-affinity, and neutralizing nanobody-targeting SVA.</i></p><p><i>• The developed nbLPB-ELISA can replace viral neutralization tests.</i></p><p><i>• The nbLPB-ELISA is suitable to evaluate protective immunity.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13492-4.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Profiling electric signals of electrogenic probiotic bacteria using self-attention analysis","authors":"Qing Chi,&nbsp;Jie Tang,&nbsp;Changqing Ji,&nbsp;Shan Chen,&nbsp;Qinquan Chen,&nbsp;Meikai Zeng,&nbsp;JiXing Cao,&nbsp;Shenxia Sun,&nbsp;Deron R. Herr,&nbsp;Qing-Gao Zhang,&nbsp;Zumin Wang,&nbsp;Chun-Ming Huang","doi":"10.1007/s00253-025-13425-1","DOIUrl":"10.1007/s00253-025-13425-1","url":null,"abstract":"<p>We fabricated a self-assembled electric circuit to detect the electrical signals produced by two electrogenic probiotic bacteria [<i>Leuconostoc mesenteroides</i> (<i>L. mesenteroides</i>) and <i>Lactococcus lactis</i> (<i>L. lactis</i>)] on chicken egg chorioallantoic membranes as well as in the intestine lumen of mice. Inoculation of <i>L. mesenteroides</i> or <i>L. lactis</i> plus glucose onto a ferrozine assay triggered the reduction of ferric ions to ferrous ions and the formation of ferrozine complexes, indicating the bacterial electron production. In the presence of glucose, <i>L. lactis</i> yielded higher electricity, measured by voltage changes, than <i>L. mesenteroides </i>in vitro. The spectra of the electrical signals generated by these two probiotic bacteria were highly distinguishable. We evaluated the importance of these differences with the application of a self-attention mechanism, a deep learning-based module, revealing several unique signals in the electrical spectra of <i>L. mesenteroides</i> as well as <i>L. lactis</i> bacteria. The specific electrical spectrum for each probiotic bacterium provided a dynamic signature for evaluation of the efficacy of various therapies using probiotics, antibiotics, and fecal microbiota transplantation in the future.</p><p>• <i> The electrical signals produced by probiotic bacteria L. mesenteroides and L. lactis on chicken egg chorioallantoic membranes and in the mouse intestine lumen were detectable.</i></p><p>• <i> In the presence of glucose, L. lactis yielded higher electricity than L. mesenteroides in vitro. Furthermore, the electrical spectra generated by these two bacteria were different.</i></p><p>• <i> The importance of these differences with the application of a self-attention mechanism revealed several unique signals in the electrical spectra.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13425-1.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143861300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a highly efficient base editing system for Lactobacilli to improve probiotics and dissect essential functions","authors":"Hitoshi Mitsunobu,&nbsp;Yudai Kita,&nbsp;Yumiko Nambu-Nishida,&nbsp;Shoko Miyazaki,&nbsp;Kensuke Nakajima,&nbsp;Ken-ichiro Taoka,&nbsp;Akihiko Kondo,&nbsp;Keiji Nishida","doi":"10.1007/s00253-025-13489-z","DOIUrl":"10.1007/s00253-025-13489-z","url":null,"abstract":"<p><i>Lactobacilli</i> play essential roles in the food industry and have a significant potential as probiotics and therapeutic agents. Genomic and genetic information has increasingly accumulated and been linked to their various functions, to which transgenic approaches are being performed to verify crucial genes. In order to reasonably develop more useful strains, beneficial traits need to be introduced into any given strains and enhanced or combined based on such genotype characterization. However, for practical use as probiotics or foods, organisms with transgene are hardly acceptable. Here, we have introduced the base editing Target-AID system specifically for <i>Lactobacilli</i>, enabling precise installation of point mutations without donor DNA and at multiple genomic loci simultaneously. <i>Lactiplantibacillus plantarum</i> has been successfully engineered to reduce production of imidazole propionate, which has been reported to be associated with type 2 diabetes by impairing glucose tolerance and insulin signaling. Additionally, this system enabled transient knock-out of an essential gene, such as one involved in cell division, resulting in severe filamentous cell phenotype. This demonstrates Target-AID is a promising genetic tool for <i>Lactobacilli</i> and can accelerate both applied and fundamental research.</p><p><i>• Efficient and multiplexable cytosine base editing established in Lactobacilli.</i></p><p><i>• Edited Lactobacillus reducing imidazole propionate associated with the risk of type 2 diabetes.</i></p><p><i>• Transient knock-out and dissection of an essential gene function.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13489-z.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143856414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering yeast for tailored fatty acid profiles","authors":"Simon Kobalter,&nbsp;Tamara Wriessnegger,&nbsp;Harald Pichler","doi":"10.1007/s00253-025-13487-1","DOIUrl":"10.1007/s00253-025-13487-1","url":null,"abstract":"<p>The demand for sustainable and eco-friendly alternatives to fossil and plant oil-derived chemicals has spurred interest in microbial production of lipids, particularly triacylglycerols, fatty acids, and their derivatives. Yeasts are promising platforms for synthesizing these compounds due to their high lipid accumulation capabilities, robust growth, and <i>g</i>enerally <i>r</i>ecognized <i>a</i>s <i>s</i>afe (GRAS) status. There is vast interest in fatty acid and triacylglycerol products with tailored fatty acid chain lengths and compositions, such as polyunsaturated fatty acids and substitutes for cocoa butter and palm oil. However, microbes naturally produce a limited set of mostly long-chain fatty acids, necessitating the development of microbial cell factories with customized fatty acid profiles. This review explores the capabilities of key enzymes involved in fatty acid and triacylglycerol synthesis, including fatty acid synthases, desaturases, elongases, and acyltransferases. It discusses factors influencing fatty acid composition and presents engineering strategies to enhance fatty acid synthesis. Specifically, we highlight successful engineering approaches to modify fatty acid profiles in triacylglycerols and produce tailored fatty acids, and we offer recommendations for host selection to streamline engineering efforts.</p><p>• <i>Detailed overview on all basic aspects of fatty acid metabolism in yeast</i></p><p>• <i>Comprehensive description of fatty acid profile tailoring in yeast</i></p><p>• <i>Extensive summary of applying tailored fatty acid profiles in production processes</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13487-1.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143861298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of exporter proteins and their engineering on the productivity of Corynebacterium","authors":"Keita Kinose,&nbsp;Keiko Shinoda,&nbsp;Hisashi Kawasaki","doi":"10.1007/s00253-025-13479-1","DOIUrl":"10.1007/s00253-025-13479-1","url":null,"abstract":"<p>Enhancing product efflux is crucial in improving fermentative bioproduction. Despite advancements in metabolic engineering guided by the design-build-test-learn cycle, membrane transport engineering of product efflux remains underdeveloped, limiting the efficient production of target chemicals. This review explores the historical findings on product efflux, regardless of passive or active transport, in fermentation engineering, focusing on <i>Corynebacterium</i> species, and highlights the potential of multidrug transporters as valuable screening sources for efflux improvement. Furthermore, the review emphasizes the importance of understanding the machinery of efflux transporters to optimize their functionality. Molecular dynamics simulations are a promising tool for exploring novel strategies to advance fermentation-related processes. These insights provide a framework for overcoming current challenges in membrane transport engineering of product efflux and improving industrial-scale bioproduction.</p><p>\u0000<i>• Review of strategies to enhance product efflux in Corynebacterium species.</i></p><p>\u0000<i>• Multidrug transporters are key tools for optimizing metabolite efflux.</i></p><p>\u0000<i>• Efflux transporter mechanisms analyzed to improve microbial productivity.</i></p><p>\u0000<i>• Molecular dynamics simulations employed for understanding transporter mechanisms.</i>\u0000</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13479-1.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143861386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 microbiome-derived metabolite trimethylamine N-oxide is associated with chronic kidney disease risk","authors":"Junyi Jiang,&nbsp;Peng Zhu,&nbsp;Xiaoying Ding,&nbsp;Li Zhou,&nbsp;Xiaoqiang Li,&nbsp;Yuyan Lei,&nbsp;Hao Wang,&nbsp;LuLu Chen,&nbsp;Xiang Li,&nbsp;Yunzhou Fei,&nbsp;Dongsheng Ouyang,&nbsp;Xiaohui Li,&nbsp;Wei Zhang","doi":"10.1007/s00253-025-13481-7","DOIUrl":"10.1007/s00253-025-13481-7","url":null,"abstract":"<p>Previous studies have established a correlation between the microbiome-derived metabolite trimethylamine <i>N</i>-oxide (TMAO) and decreased renal function, but with great heterogeneity. Moreover, population-based evidence remains scarce, particularly in Chinese populations. We designed a meta-analysis and a population-based cross-sectional study in China to examine the associations between TMAO and chronic kidney disease (CKD). In meta-analysis, among 2125 pooled subjects with 1240 controls and 885 CKD patients, a significant association was observed between TMAO and CKD, with a standardized mean difference of − 0.93 (95% confidence interval: − 1.11, − 0.75). Meta-regression analysis identified gender, age, and body mass index (BMI) as significant heterogeneity factors. In our population-based study of 5584 subjects with an estimated glomerular filtration rate (eGFR) ≥ 60 mL/min/1.73 m² from Sijing community, 100 developed CKD in 2 years. We matched 195 controls by age and gender from the 5484 non-CKD subjects. Male subjects and alcohol consumers exhibited a lower risk of CKD with adjusted odds ratio (OR) of 0.471 (<i>P</i> &lt; 0.05) and 0.320 (<i>P</i> &lt; 0.05), respectively. When comparing subjects in the lowest tertile of TMAO, adjusted OR reached to 1.243 (<i>P</i> &gt; 0.05) for those in the middle and 2.123 (<i>P</i> &lt; 0.05) in the highest tertile (<i>P</i> for trend &lt; 0.05). TMAO demonstrated a moderate capacity to distinguish CKD from non-CKD subjects (AUC = 0.614, <i>P</i> &lt; 0.01). Our findings indicate TMAO is significantly associated with the risk of CKD, and suggest age, gender, and BMI may confound the relationship between TMAO and CKD.</p><p><i>• Subjects with elevated TMAO levels have an increased risk of CKD.</i></p><p><i>• TMAO demonstrates a moderate capacity to distinguish CKD from non-CKD cases.</i></p><p><i>• Age, gender and BMI may confound the relationship between TMAO and CKD.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13481-7.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143856413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mesoporous silica and vegetal extracts combined as sustainable stone heritage protection against biodeterioration","authors":"Andrea Campostrini,&nbsp;Agustí Sala-Luis,&nbsp;Pilar Bosch-Roig,&nbsp;Elena Ghedini,&nbsp;Michela Signoretto,&nbsp;Federica Menegazzo","doi":"10.1007/s00253-025-13475-5","DOIUrl":"10.1007/s00253-025-13475-5","url":null,"abstract":"<p>Since biodeterioration is considered one of the main issues related to the conservation of cultural heritage stone materials, an investigation was conducted into preventive sustainable antimicrobial alternatives to protect the stone surfaces. The study focuses on using MCM-41 mesoporous silica particles and vegetal extracts: the mesoporous materials act as nanocontainers encapsulating the extracts, which instead serve as green antimicrobic compounds to inhibit microbiological proliferation. In this way, the antimicrobial features of the extracts are sustained for a more extended period, reducing the evaporation rate and diminishing the quantity required; the amount necessary to achieve the minimum inhibitory concentration was reduced due to the decrease in evaporation. Moreover, since the MCM-41 can host a higher quantity of product than is necessary to exert the antimicrobial effect, the duration of activity is further prolonged, releasing the extracts over time. Specifically, the mesoporous particles were impregnated with the vegetal extract of limonene and the essential oils of thyme and oregano. In vitro microbiological tests were conducted on two fungi (i.e., <i>Aspergillus tubingensis</i> and <i>Penicillium chrysogenum</i>), taken as model microorganisms from real-case scenarios. A combination of mesoporous silica and vegetal extracts was employed to develop a protective coating for stone surfaces, and tests were conducted on marble mock-ups. The promising synergic results show that this system could be of interest for preventing microbiological growth over stone surfaces, avoiding a visible aesthetic impact, being non-toxic for the environment or the operator, and preventing the extract from evaporating but holding it for a controlled release.</p><p>• <i>Green antimicrobial system using porous silica as nanocontainer for plant extracts</i></p><p>• <i>Encapsulated vegetal extracts to inhibit microbial growth on stone surfaces</i></p><p>• <i>Stable and efficient coating against fungal species in vitro and on marble mock-up</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13475-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143861301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biosynthesis of ergothioneine: current state, achievements, and perspectives","authors":"Shun Sato,&nbsp;Azusa Saika,&nbsp;Tatsuyuki Koshiyama,&nbsp;Yukihiro Higashiyama,&nbsp;Tokuma Fukuoka,&nbsp;Tomotake Morita","doi":"10.1007/s00253-025-13476-4","DOIUrl":"10.1007/s00253-025-13476-4","url":null,"abstract":"<p>Ergothioneine (EGT) is a derivative of the amino acid L-histidine that is well known for its strong antioxidant properties. Recent studies on the functional characterization of EGT in both in vivo and in vitro systems have demonstrated its potential applications in pharmaceuticals, food, and cosmetics. The growing demand for EGT in novel applications necessitates the development of safe and cost-effective mass production technologies. Consequently, microbial fermentation for EGT biosynthesis has attracted significant attention. This review focuses on the biosynthesis of EGT via microbial fermentation, explores its biosynthetic mechanisms, and summarizes the latest advancements for industrial EGT production using engineered microbial strains.</p><p>• <i>Ergothioneine (EGT) is an L-histidine derivative with strong antioxidant property.</i></p><p>• <i>Recent studies have revealed certain groups of microbes produce EGT naturally.</i></p><p>• <i>Superior EGT producers by genetic modification have been created.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13476-4.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143821966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harnessing the power of biosensors for environmental monitoring of pesticides in water","authors":"Filipa Mendes,&nbsp;Beatriz O. Machado,&nbsp;Bruno B. Castro,&nbsp;Maria João Sousa,&nbsp;Susana R. Chaves","doi":"10.1007/s00253-025-13461-x","DOIUrl":"10.1007/s00253-025-13461-x","url":null,"abstract":"<p>The current strong reliance on synthetic chemicals, namely pesticides, is far from environmentally sustainable. These xenobiotics contribute significantly to global change and to the current biodiversity crisis, but have been overlooked when compared to other agents (e.g., climate change). Aquatic ecosystems are particularly vulnerable to pesticides, making monitoring programs essential to preserve ecosystem health, safeguard biodiversity, ensure water quality, and mitigate potential human health risks associated with contaminated water sources. Biosensors show great potential as time/cost-effective and disposable systems for the high-throughput detection (and quantification) of these pollutants. In this mini-review, we provide an overview of biosensors specifically developed for environmental water monitoring, covering different pesticide classes (and active ingredients), and types of biosensors (according to the bio-recognition element) and transducers, as well as the nature of sample matrices analyzed. We highlight the variety of biosensors that have been developed and successfully applied to detection of pesticides in aqueous samples, including enzymatic biosensors, immunosensors, aptasensors, and whole cell–based biosensors. While most biosensors have been designed to detect insecticides, expanding their compound target range could significantly streamline monitoring of environmental contaminants. Despite limitations related to stability, reproducibility, and interference from environmental factors, biosensors represent a promising and sustainable technology for pesticide monitoring in the aquatic environments, offering sensitivity and specificity, as well as portability and real-time results. We propose that biosensors would be most effective as an initial screening step in a tiered assessment, complementing conventional methods.</p><p>• <i>Pesticides harm aquatic ecosystems and biodiversity, requiring better monitoring</i></p><p>• <i>Biosensors offer cost-effective solutions to detect pesticides in water samples</i></p><p>• <i>Biosensors complement conventional methods as a sustainable tool for initial screens</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13461-x.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143821967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mycotoxin-free Aspergillus oryzae strain lineage for alternative and novel protein production at industrial scale","authors":"Jan Lehmbeck,&nbsp;Birgitte Andersen,&nbsp;Javier Sáez-Sáez,&nbsp;Jens Christian Frisvad,&nbsp;José Arnau","doi":"10.1007/s00253-025-13482-6","DOIUrl":"10.1007/s00253-025-13482-6","url":null,"abstract":"<p>Advanced industrial strains of <i>Aspergillus oryzae</i> have been used for decades for the production of recombinant proteins including food and feed enzymes at large scale. The <i>A. oryzae</i> strain lineage evaluated in this review derives from the proprietary Novozymes (now Novonesis) strain collection. <i>A. oryzae</i> wild-type strains have the potential to produce three different mycotoxins (aflatoxins (AFL), cyclopiazonic acid (CPA), and 3-nitropropionic acid (3-NPA)). Here, we review the work originally performed at Novozymes to identify a strain (BECh1) that contained a large chromosomal deletion comprising both AFL and CPA gene clusters, significantly improving the safety of the lineage. The description of the deleted region is presented here. As the genetic basis for 3-NPA biosynthesis was recently revealed, we describe here that this <i>A. oryzae</i> lineage contains an additional large deletion that encompasses the 3-NPA biosynthetic genes <i>npaA</i> and <i>npaB</i>, thereby rendering the strains unable to produce any mycotoxin. Further strain development has resulted in strains devoid of penicillin production by inactivation of the penicillin G gene cluster (<i>penG</i>). This strain lineage represents the first example of mycotoxin-free <i>A. oryzae</i> for production of recombinant (alternative) novel food proteins. Recently, bovine beta-lactoglobulin made using a strain of this lineage has received GRAS status and can be commercialized for use in food in the USA. With its history of safe use in food and feed, the lack of toxigenic potential and the ability to differentiate strains with modern technologies, this <i>A. oryzae</i> strain can be considered safe as other organisms with a Quality Presumption of Safety (QPS) status in Europe. QPS is not applicable to filamentous fungi and only granted at the species level to bacteria and a few yeast species. We suggest modernizing the QPS concept to become strain rather than species specific and present arguments to qualify this strain lineage as QPS or QPS-like.</p><p><i>• The biosynthetic genes for 3-nitropropionic acid have been recently characterized.</i></p><p><i>• An industrial strain lineage for food protein production lacks all known mycotoxin genes.</i></p><p><i>• Proteins produced in this mycotoxin-free lineage should entail lower regulatory requirements.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13482-6.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143824587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}