Microbial Biotechnology最新文献

One-Step RAA and CRISPR-Cas13a Method for Detecting Influenza B Virus

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-15 DOI: 10.1111/1751-7915.70144

Xinling Zhang, Shiyu Chen, Juezhuo Li, Dong-ang Liu, Jianxiu Lai, Xiangquan Song, Ruiyao Hu, Yuting Qiu, Keyi Chen, Yue Xu, Xiaoping Li

{"title":"One-Step RAA and CRISPR-Cas13a Method for Detecting Influenza B Virus","authors":"Xinling Zhang, Shiyu Chen, Juezhuo Li, Dong-ang Liu, Jianxiu Lai, Xiangquan Song, Ruiyao Hu, Yuting Qiu, Keyi Chen, Yue Xu, Xiaoping Li","doi":"10.1111/1751-7915.70144","DOIUrl":"https://doi.org/10.1111/1751-7915.70144","url":null,"abstract":"We developed a sensitive and specific method based on recombinase-aided amplification (RAA) and clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 13a (Cas13a). This method, named CRISPR-based Rapid and Efficient Test (CRISPRET), is designed for the early diagnosis of Influenza B (FluB) with the aim of shortening its transmission chain. We identified conserved regions in the Influenza B Virus (IBV) NS gene and designed forward and reverse primers along with crRNAs. We then established and optimised the reaction system, and Nucleic Acid Positive Reference Materials of IBV were used to evaluate the detection limit (DL) of CRISPRET. Additionally, we collected 257 clinical samples, comprising 127 samples from patients with IBV infection and 130 samples from healthy individuals, and subjected them to dual detection using CRISPRET and qPCR to evaluate the positive predictive value (PPV), negative predictive value (NPV), sensitivity and specificity of CRISPRET. We designed one forward primer, two reverse primers, and two crRNAs to establish and optimise the CRISPR ET. The method demonstrated the DL of 500 copies·μL−1 when assisted by appropriate equipment. Despite requiring auxiliary equipment and a 30-min reaction, the CRISPR ET method enables the detection of IBV nucleic acid within approximately the first 5 min, achieving high sensitivity (100%), specificity (97.69%), PPV (97.69%) and NPV (100%), with a concordance rate of 98.83% to qPCR. CRISPRET offers a simple, field-applicable, one-step method for the rapid detection of IBV. It has strong potential for field-testing applications and intelligent integration into existing diagnostic systems.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70144","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143831257","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}

引用次数: 0

Unlocking the Microbial World: A Global Initiative for Education and Sustainability

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-07 DOI: 10.1111/1751-7915.70124

Juan L. Ramos, Rup Lal, Francisca Colom, Max Chavarria, Wei Huang, Yun Wang, Zulema Udaondo, Kenneth N. Timmis

引用次数: 0

Monkeypox Virus: WHO's Second Public Health Emergency of International Concern Within 2 Years

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-07 DOI: 10.1111/1751-7915.70142

Harald Brüssow

{"title":"Monkeypox Virus: WHO's Second Public Health Emergency of International Concern Within 2 Years","authors":"Harald Brüssow","doi":"10.1111/1751-7915.70142","DOIUrl":"https://doi.org/10.1111/1751-7915.70142","url":null,"abstract":"An upsurge of monkeypox disease (mpox) cases with clade I virus in Central Africa led WHO to declare a Public Health Emergency of International Concern for a second time shortly after the worldwide clade II mpox epidemic in 2022/3 among homosexual men. In the Democratic Republic of Congo (DRC), the annual incidence of clade I mpox, transmitted mostly from animal sources to children, increased 20-fold between 1980 and 2007; 60,000 mpox cases occurred between 2010 and 2023. The incidence again doubled between 2023 and 2024, showing a case fatality rate of 3.3%. A new clade Ib virus was detected in 2024 in eastern DRC where mostly adults were infected by heterosexual contact. Ib was recently introduced and showed a mutation spectrum of human-to-human transmission. Asymptomatic mpox infections, the release of infectious virus before symptom onset in a subgroup of cases, and superspreaders complicate containment measures during the 2022 epidemic. Isolation of cases until two consecutive negative PCR tests was recommended but necessitates cheap and rapid diagnostic tests which are in development. Sexual behavioural changes during the 2022 epidemic have contributed more to the curbing of the epidemic than vaccination. The smallpox vaccine Dryvax protected children exposed to clade I mpox in DRC in the 1980s. The attenuated third-generation smallpox Modified Vaccinia Ankara (MVA) vaccines and derivatives showed robust protection against clade IIb mpox during the 2022/3 epidemic in various study formats. Vaccine efficacy exceeding 75% was reported after two doses. mRNA in lipid-nanoparticle encoding surface proteins from extracellular enveloped and intracellular mature virions of monkeypox virus (MPXV) induced humoral and cellular immune responses that protected macaques against mpox disease with clade I and II viruses better than MVA. Only mixtures of monoclonal antibodies protected mice from mpox. The antiviral tecovirimat showed no efficacy in two clinical trials against clade I and II mpox.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70142","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143786971","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}

引用次数: 0

Metabolic Engineering of a Serotonin Overproducing Saccharomyces cerevisiae Strain

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-05 DOI: 10.1111/1751-7915.70140

Andrés Planells-Cárcel, Elena Valera-García, Guillermo Quintas, José Luis Martínez, Sara Muñiz-Calvo, José Manuel Guillamón

{"title":"Metabolic Engineering of a Serotonin Overproducing Saccharomyces cerevisiae Strain","authors":"Andrés Planells-Cárcel, Elena Valera-García, Guillermo Quintas, José Luis Martínez, Sara Muñiz-Calvo, José Manuel Guillamón","doi":"10.1111/1751-7915.70140","DOIUrl":"https://doi.org/10.1111/1751-7915.70140","url":null,"abstract":"The EU Green Deal prioritises the transformation of the chemical industry to a more environmentally sustainable model. This involves using microorganisms, such as Saccharomyces cerevisiae, to produce molecules more sustainably through biotechnological approaches. In this study, we demonstrate an example of serotonin production using S. cerevisiae as a cell factory, along with its optimisation and upscaling. To achieve this, we introduced two heterologous genes, the combination of tryptophan decarboxylase from Clostridium sporogenes (CsTDC) and tryptamine 5-hydroxylase from Oryza sativa (OsT5H), to complete the serotonin biosynthetic pathway using L-tryptophan (L-TRP) as a precursor. By modifying ARO4 to a feedback-resistant version (ARO4*), the flux of the shikimate pathway was significantly increased and serotonin production was achieved at levels up to 120 mg/L directly from the glucose source. After a medium optimisation, a final concentration of 80 g/L glucose and 300 mg/L of nitrogen resulted in better conditions for increasing serotonin titres. Using this medium in a 1 L bioreactor fermentation resulted in approximately 250 mg/L of serotonin. A targeted metabolomic study of the bioreactor growth medium identified potential bottlenecks in the serotonin-overproducing strain and future targets for increasing its titre. We have constructed a strain of S. cerevisiae that represents the first steps towards feasible industrial production of serotonin using a sustainable and environmentally friendly approach, paving the way for the development of similar biotechnological strategies in the future.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70140","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143778233","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}

引用次数: 0

Gamifying Science—Novel Strategies for Bringing Microbiology to the Classroom 科学游戏化--将微生物学带入课堂的新策略

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-05 DOI: 10.1111/1751-7915.70141

Carlos del Cerro-Sánchez, Vanesa Amarelle, Raúl Platero, Danilo Pérez-Pantoja, Daniela Wicki-Emmenegger, Max Chavarría, Gonzalo Durante-Rodríguez

引用次数: 0

Exploring Yeast's Energy Dynamics: The General Stress Response Lowers Maintenance Energy Requirement

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-03 DOI: 10.1111/1751-7915.70126

Nuran Temelli, Simon van den Akker, Ruud A. Weusthuis, Markus M. M. Bisschops

{"title":"Exploring Yeast's Energy Dynamics: The General Stress Response Lowers Maintenance Energy Requirement","authors":"Nuran Temelli, Simon van den Akker, Ruud A. Weusthuis, Markus M. M. Bisschops","doi":"10.1111/1751-7915.70126","DOIUrl":"https://doi.org/10.1111/1751-7915.70126","url":null,"abstract":"In many microbial biotechnology processes, biomass itself is not the product of interest, but rather targeted chemicals or proteins. In these processes, growth should be limited to direct more substrate to product and increase process yields. Under growth-limiting conditions, such as nutrient limitation, microorganisms, including the yeast Saccharomyces cerevisiae, activate a general stress response (GSR). Different hypotheses have been formulated for this activation, including a preparatory role for future stresses or a role in cellular protein density. Here we tested a third hypothesis: the GSR reduces the energy needed to maintain cellular homeostasis, also known as the maintenance energy requirement (MER). The impact of GSR on MER was investigated by assessing the effect of the absence of its key regulators, Msn2 and Msn4, on energy-substrate distribution and stress resistance. Chemostat and fed-batch cultures revealed significant increases in MER of up to 85% in the deletion strain compared to the parental strain. In contrast, maximal biomass yields, growth rates and morphology were unaffected. Our insights highlight an additional role of the GSR, namely saving cellular energy. As the MER is a key determinant of product yields and in process design, especially in low growth processes, our findings can help to optimise microbial bioprocesses.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70126","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770093","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}

引用次数: 0

De Novo Biosynthesis of Antidepressant Psilocybin in Escherichia coli

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-03 DOI: 10.1111/1751-7915.70135

Zhangrao Huang, Yongpeng Yao, Rouyu Di, JianChao Zhang, Yuanyuan Pan, Gang Liu

{"title":"De Novo Biosynthesis of Antidepressant Psilocybin in Escherichia coli","authors":"Zhangrao Huang, Yongpeng Yao, Rouyu Di, JianChao Zhang, Yuanyuan Pan, Gang Liu","doi":"10.1111/1751-7915.70135","DOIUrl":"https://doi.org/10.1111/1751-7915.70135","url":null,"abstract":"Psilocybin, a tryptamine-derived alkaloid, has been granted Breakthrough Therapy designation by the U.S. FDA for treatment-resistant depression, underscoring its clinical importance. Therefore, sustainable and economic production is urgently needed. Manufacturing of psilocybin in Escherichia coli has drawn great attention. However, due to the low expression and activity of the eukaryotic cytochrome P450 enzyme PsiH in the psilocybin biosynthetic pathway, de novo synthesis of psilocybin in prokaryotic cells has been hampered. To overcome this dilemma, we herein demonstrated de novo synthesis of psilocybin in E. coli by constructing PsiH variants with N-terminal domain modifications and expressing the entire biosynthetic pathway at a concordantly low temperature. Improving the supply of precursor and engineering the P450 electron transfer chain resulted in a 33-fold increase in the titre of norbaeocystin (105.3 mg/L), a key intermediate of psilocybin biosynthesis, and a 17-fold increase in the titre of psilocybin (14 mg/L). Further enhancement of psilocybin production was achieved by converting norbaeocystin to psilocybin by overexpressing an extra copy of the methyltransferase gene psiM. Finally, 79.4 mg/L of psilocybin was produced by optimising flask fermentation conditions, a 100-fold improvement over the starting strain. Our work demonstrates the successful fungal P450 engineering to improve the catalytic activity in E. coli and will advance the sustainable production of the important antidepressant psilocybin in prokaryotic microbial cells.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70135","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143761978","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}

引用次数: 0

Variation of Meat Quality and Relationship to Gut Microbiota Among Different Pig Breeds

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-03 DOI: 10.1111/1751-7915.70139

Qifan Zhang, Man Du, Siyu Wei, Luoyi Zhu, Rong Yan, Mingliang Jin, Yizhen Wang

{"title":"Variation of Meat Quality and Relationship to Gut Microbiota Among Different Pig Breeds","authors":"Qifan Zhang, Man Du, Siyu Wei, Luoyi Zhu, Rong Yan, Mingliang Jin, Yizhen Wang","doi":"10.1111/1751-7915.70139","DOIUrl":"https://doi.org/10.1111/1751-7915.70139","url":null,"abstract":"Meat production is of great importance to the world's food supply and economic development, and meat quality determines the purchasing desire of consumers. Recent studies show intestinal microorganisms are involved in several physiological functions of the host and therefore are likely to regulate meat quality. This study aimed to compare the carcass performance, meat quality traits and serum parameters of three different pig breeds, Jinhua (JH) pigs (n = 8), Duroc × Berkshire × Jiaxinghei (DBJ) pigs (n = 8), Duroc × Landrace × Yorkshire (DLY) pigs (n = 8) and to investigate a possible relationship between gut microbiota composition and these traits. Meat quality results showed that compared with DLY pigs, JH pigs had lower water loss and shear force in the longissimus dorsi muscle, and higher intramuscular fat content and inosine monophosphate content were observed in JH pigs. Serum biochemical indicators showed the content of nonesterified fatty acid in JH pigs was lowest. Furthermore, the gut microbiota analysis indicated that JH pigs harboured more abundant Lachnospiraceae, Prevotellaceae NK3B31 and Marvinbryantia. Spearman correlation analysis showed that unidentified genus of family Lachnospiraceae, genus Prevotella and genus Alloprevotella were positively correlated with IMF content and marbling score in the longissimus dorsi muscle of pigs. In conclusion, our results indicated the quality of JH pork was superior to DBJ and DLY pigs, and the difference in meat qualities was related to the abundance of fibre-degrading bacteria. Our study provides insight into further understanding of the relationship between microbiota and meat quality, nutrient metabolism and fat deposition, which is critical to the pork industry and swine intestinal health.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70139","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770095","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}

引用次数: 0

Unravelling the Glycan Code: Molecular Dynamics and Quantum Chemistry Reveal How O-Glycan Functional Groups Govern OgpA Selectivity in Mucin Degradation by Akkermansia muciniphila

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-03 DOI: 10.1111/1751-7915.70091

Mohammad Khavani, Aliyeh Mehranfar, Mohammad R. K. Mofrad

{"title":"Unravelling the Glycan Code: Molecular Dynamics and Quantum Chemistry Reveal How O-Glycan Functional Groups Govern OgpA Selectivity in Mucin Degradation by Akkermansia muciniphila","authors":"Mohammad Khavani, Aliyeh Mehranfar, Mohammad R. K. Mofrad","doi":"10.1111/1751-7915.70091","DOIUrl":"https://doi.org/10.1111/1751-7915.70091","url":null,"abstract":"Mucins, heavily O-glycosylated glycoproteins, are a key component of mucus, and certain gut microbiota, including Akkermansia muciniphila, can utilise mucin glycans as a carbon source. Akkermansia muciniphila produces the O-glycopeptidase enzyme OgpA, which cleaves peptide bonds at the N-terminus of serine (Ser) or threonine (Thr) residues carrying O-glycan substitutions, with selectivity influenced by the O-glycan functional groups. Using molecular dynamics (MD) simulations and quantum chemistry calculations, we explored how different O-glycan groups affect OgpA's selectivity. Our results show that peptides bind to the enzyme via hydrogen bonds, π–π interactions, van der Waals forces and electrostatic interactions, with key residues, including Tyr90, Val138, Gly176, Tyr210 and Glu91, playing important roles. The primary determinant of selectivity is the interaction between the peptide's functional group and the enzyme's binding cavity, while peptide–enzyme interface interactions are secondary. Quantum chemistry calculations reveal that OgpA prefers peptides with a lower electrophilic character. This study provides new insights into mucin degradation by gut microbiota enzymes, advancing our understanding of this critical biological process.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70091","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770092","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}

引用次数: 0

Genetic Dissection of Cyclic di-GMP Signalling in Pseudomonas aeruginosa via Systematic Diguanylate Cyclase Disruption

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2025-04-02 DOI: 10.1111/1751-7915.70137

Román A. Martino, Daniel C. Volke, Albano H. Tenaglia, Paula M. Tribelli, Pablo I. Nikel, Andrea M. Smania

{"title":"Genetic Dissection of Cyclic di-GMP Signalling in Pseudomonas aeruginosa via Systematic Diguanylate Cyclase Disruption","authors":"Román A. Martino, Daniel C. Volke, Albano H. Tenaglia, Paula M. Tribelli, Pablo I. Nikel, Andrea M. Smania","doi":"10.1111/1751-7915.70137","DOIUrl":"https://doi.org/10.1111/1751-7915.70137","url":null,"abstract":"The second messenger bis-(3′ → 5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) governs adaptive responses in the opportunistic pathogen Pseudomonas aeruginosa, including biofilm formation and the transition from acute to chronic infections. Understanding the intricate c-di-GMP signalling network remains challenging due to the overlapping activities of numerous diguanylate cyclases (DGCs). In this study, we employed a CRISPR-based multiplex genome-editing tool to disrupt all 32 GGDEF domain-containing proteins (GCPs) implicated in c-di-GMP signalling in P. aeruginosa PA14. Phenotypic and physiological analyses revealed that the resulting mutant was unable to form biofilms and had attenuated virulence. Residual c-di-GMP levels were still detected despite the extensive GCP disruption, underscoring the robustness of this regulatory network. Taken together, these findings provide insights into the complex c-di-GMP metabolism and showcase the importance of functional overlapping in bacterial signalling. Moreover, our approach overcomes the native redundancy in c-di-GMP synthesis, providing a framework to dissect individual DGC functions and paving the way for targeted strategies to address bacterial adaptation and pathogenesis.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 4","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70137","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143749548","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}

引用次数: 0