Isabel De La Torre, Miguel G. Acedos, Juan J. Cestero, Jorge Barriuso, José L. García
{"title":"Engineering Xylose Isomerase and Reductase Pathways in Yarrowia lipolytica for Efficient Lipid Production","authors":"Isabel De La Torre, Miguel G. Acedos, Juan J. Cestero, Jorge Barriuso, José L. García","doi":"10.1111/1751-7915.70127","DOIUrl":"https://doi.org/10.1111/1751-7915.70127","url":null,"abstract":"<p>Xylose is a common monosaccharide in lignocellulosic residues that <i>Yarrowia lipolytica</i> cannot naturally metabolise for lipid production and therefore, heterologous xylose metabolic pathways must be engineered in this yeast to facilitate its consumption. We have compared the metabolic efficiency of two xylose metabolic pathways by developing three recombinant <i>Y. lipolytica</i> strains: one harbouring a xylose reductase pathway, one with a xylose isomerase pathway, and one combining both pathways, and the strains were tested for xylose consumption and lipid production at different scales. The recombinant strain with the reductase pathway that was directly isolated in selective xylose medium showed the highest lipid yield, producing up to 12.8 g/L of lipids, or 43% of the biomass dry weight, without requiring any other xylose consumption adaptive evolution process. This strain achieved a lipid yield of 0.13 g lipids/g xylose, one of the highest yields in yeast reported so far using xylose as the sole carbon and energy source. Although the strain harbouring the isomerase pathway performed better under oxygen-limiting conditions and led to higher lipid intracellular accumulation, it showed a lower xylose uptake and biomass production, rendering a lower yield under non-limiting oxygen conditions. Unexpectedly, the combination of both pathways in the same strain was less effective than the use of the reductase pathway alone.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70127","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143629813","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}
Isabel De La Torre, Miguel G. Acedos, Juan J. Cestero, Jorge Barriuso, José L. García
{"title":"Engineering Xylose Isomerase and Reductase Pathways in Yarrowia lipolytica for Efficient Lipid Production","authors":"Isabel De La Torre, Miguel G. Acedos, Juan J. Cestero, Jorge Barriuso, José L. García","doi":"10.1111/1751-7915.70127","DOIUrl":"https://doi.org/10.1111/1751-7915.70127","url":null,"abstract":"<p>Xylose is a common monosaccharide in lignocellulosic residues that <i>Yarrowia lipolytica</i> cannot naturally metabolise for lipid production and therefore, heterologous xylose metabolic pathways must be engineered in this yeast to facilitate its consumption. We have compared the metabolic efficiency of two xylose metabolic pathways by developing three recombinant <i>Y. lipolytica</i> strains: one harbouring a xylose reductase pathway, one with a xylose isomerase pathway, and one combining both pathways, and the strains were tested for xylose consumption and lipid production at different scales. The recombinant strain with the reductase pathway that was directly isolated in selective xylose medium showed the highest lipid yield, producing up to 12.8 g/L of lipids, or 43% of the biomass dry weight, without requiring any other xylose consumption adaptive evolution process. This strain achieved a lipid yield of 0.13 g lipids/g xylose, one of the highest yields in yeast reported so far using xylose as the sole carbon and energy source. Although the strain harbouring the isomerase pathway performed better under oxygen-limiting conditions and led to higher lipid intracellular accumulation, it showed a lower xylose uptake and biomass production, rendering a lower yield under non-limiting oxygen conditions. Unexpectedly, the combination of both pathways in the same strain was less effective than the use of the reductase pathway alone.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70127","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143629812","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}
Wan-Qi Qin, Yi-Fan Liu, Jin-Feng Liu, Lei Zhou, Shi-Zhong Yang, Ji-Dong Gu, Bo-Zhong Mu
{"title":"The Metabolic and Physiological Responses to Spaceflight of a Lipopeptide-Producing Bacillus subtilis","authors":"Wan-Qi Qin, Yi-Fan Liu, Jin-Feng Liu, Lei Zhou, Shi-Zhong Yang, Ji-Dong Gu, Bo-Zhong Mu","doi":"10.1111/1751-7915.70111","DOIUrl":"https://doi.org/10.1111/1751-7915.70111","url":null,"abstract":"<p>Outer space is an extreme environment and the survival of many microorganisms after spaceflight is well established. However, adaptations of <i>Bacillus subtilis</i> to space stress, particularly metabolism, are largely unknown. Here, we first performed a spaceflight mission of the <i>B. subtilis</i> TD7 strain and compared the spaceflight-exposed strain with the wild-type in terms of their phenotype, biofilm formation and secondary metabolism. The spaceflight-exposed strain exhibited slower growth, different morphology and decreased biofilm formation. Importantly, a decline in lipopeptide production was observed after spaceflight. Multi-omics approaches were used to uncover the molecular mechanisms underlying secondary metabolism and 997 differentially expressed genes (DEGs) were found, involving the TCA cycle, fatty acid degradation, amino acid biosynthesis and quorum sensing systems. Further analysis of 26 lipopeptide-related DEGs further elucidated the relationship between the space environment and secondary metabolism regulation. Our findings could contribute to a better understanding of the relationship between the space environment and microbial adaptation mechanisms.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602467","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}
Wan-Qi Qin, Yi-Fan Liu, Jin-Feng Liu, Lei Zhou, Shi-Zhong Yang, Ji-Dong Gu, Bo-Zhong Mu
{"title":"The Metabolic and Physiological Responses to Spaceflight of a Lipopeptide-Producing Bacillus subtilis","authors":"Wan-Qi Qin, Yi-Fan Liu, Jin-Feng Liu, Lei Zhou, Shi-Zhong Yang, Ji-Dong Gu, Bo-Zhong Mu","doi":"10.1111/1751-7915.70111","DOIUrl":"https://doi.org/10.1111/1751-7915.70111","url":null,"abstract":"<p>Outer space is an extreme environment and the survival of many microorganisms after spaceflight is well established. However, adaptations of <i>Bacillus subtilis</i> to space stress, particularly metabolism, are largely unknown. Here, we first performed a spaceflight mission of the <i>B. subtilis</i> TD7 strain and compared the spaceflight-exposed strain with the wild-type in terms of their phenotype, biofilm formation and secondary metabolism. The spaceflight-exposed strain exhibited slower growth, different morphology and decreased biofilm formation. Importantly, a decline in lipopeptide production was observed after spaceflight. Multi-omics approaches were used to uncover the molecular mechanisms underlying secondary metabolism and 997 differentially expressed genes (DEGs) were found, involving the TCA cycle, fatty acid degradation, amino acid biosynthesis and quorum sensing systems. Further analysis of 26 lipopeptide-related DEGs further elucidated the relationship between the space environment and secondary metabolism regulation. Our findings could contribute to a better understanding of the relationship between the space environment and microbial adaptation mechanisms.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602567","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}
Fan Liu, Jingwen Zhou, Jianghua Li, Jian Chen, Guocheng Du, Xinrui Zhao
{"title":"Precise Engineering and Efficient Biosynthesis of Robust and High-Activity Human Haemoglobin for Artificial Oxygen Carriers","authors":"Fan Liu, Jingwen Zhou, Jianghua Li, Jian Chen, Guocheng Du, Xinrui Zhao","doi":"10.1111/1751-7915.70128","DOIUrl":"https://doi.org/10.1111/1751-7915.70128","url":null,"abstract":"<p>Recombinant human haemoglobin (rHb) is a tetramer protein with heme as cofactors, which have extensive applications in the fields of biomaterials and biomedical therapeutics. However, due to the poor structural stability, the dissociation of heme, weak oxygen transport efficiency, and lower activity, the utilisation of rHb is severely limited in artificial oxygen carriers. Herein, based on the novel developed high-throughput screening strategies and semi-rational design, the engineered rHb mutant with strong stability and heme-binding ability was obtained. In addition, through the homology alignment and rational design, the oxygen transport capacity of rHb was significantly enhanced. Furthermore, the bottlenecks of heme supply were overcome by applying the fine-tuned heme synthesis in <i>Escherichia coli</i>. Finally, the robust and high-activity rHb mutant was synthesised and can be used as a new generation of artificial oxygen carriers.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70128","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602669","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 Legalome: Microbiology, Omics and Criminal Justice","authors":"Alan C. Logan, Pragya Mishra, Susan L. Prescott","doi":"10.1111/1751-7915.70129","DOIUrl":"https://doi.org/10.1111/1751-7915.70129","url":null,"abstract":"<p>Advances in neuromicrobiology and related omics technologies have reinforced the idea that unseen microbes play critical roles in human cognition and behaviour. Included in this research is evidence indicating that gut microbes, through direct and indirect pathways, can influence aggression, anger, irritability and antisocial behaviour. Moreover, gut microbes can manufacture chemicals that are known to compromise cognition. For example, recent court decisions in the United States and Europe acknowledge that gut microbes can produce high levels of ethanol, without consumption of alcohol by the defendants. The dismissal of driving while intoxicated charges in these cases—so-called auto-brewery syndrome—highlights the way in which microbiome knowledge will enhance the precision, objectivity and fairness of our legal systems. Here in this opinion essay, we introduce the concept of the ‘legalome’—the application of microbiome and omics science to forensic psychiatry and criminal law. We argue that the rapid pace of microbial discoveries, including those that challenge ideas of free will and moral responsibility, will necessitate a reconsideration of traditional legal doctrines and justifications of retributive punishment. The implications extend beyond the courtroom, challenging us to reconsider how environmental factors—from diet to socioeconomic conditions—might shape preventative and rehabilitative efforts through their effects on the microbiome.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70129","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143595394","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}
Fan Liu, Jingwen Zhou, Jianghua Li, Jian Chen, Guocheng Du, Xinrui Zhao
{"title":"Precise Engineering and Efficient Biosynthesis of Robust and High-Activity Human Haemoglobin for Artificial Oxygen Carriers","authors":"Fan Liu, Jingwen Zhou, Jianghua Li, Jian Chen, Guocheng Du, Xinrui Zhao","doi":"10.1111/1751-7915.70128","DOIUrl":"https://doi.org/10.1111/1751-7915.70128","url":null,"abstract":"<p>Recombinant human haemoglobin (rHb) is a tetramer protein with heme as cofactors, which have extensive applications in the fields of biomaterials and biomedical therapeutics. However, due to the poor structural stability, the dissociation of heme, weak oxygen transport efficiency, and lower activity, the utilisation of rHb is severely limited in artificial oxygen carriers. Herein, based on the novel developed high-throughput screening strategies and semi-rational design, the engineered rHb mutant with strong stability and heme-binding ability was obtained. In addition, through the homology alignment and rational design, the oxygen transport capacity of rHb was significantly enhanced. Furthermore, the bottlenecks of heme supply were overcome by applying the fine-tuned heme synthesis in <i>Escherichia coli</i>. Finally, the robust and high-activity rHb mutant was synthesised and can be used as a new generation of artificial oxygen carriers.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70128","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602670","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 Legalome: Microbiology, Omics and Criminal Justice","authors":"Alan C. Logan, Pragya Mishra, Susan L. Prescott","doi":"10.1111/1751-7915.70129","DOIUrl":"https://doi.org/10.1111/1751-7915.70129","url":null,"abstract":"<p>Advances in neuromicrobiology and related omics technologies have reinforced the idea that unseen microbes play critical roles in human cognition and behaviour. Included in this research is evidence indicating that gut microbes, through direct and indirect pathways, can influence aggression, anger, irritability and antisocial behaviour. Moreover, gut microbes can manufacture chemicals that are known to compromise cognition. For example, recent court decisions in the United States and Europe acknowledge that gut microbes can produce high levels of ethanol, without consumption of alcohol by the defendants. The dismissal of driving while intoxicated charges in these cases—so-called auto-brewery syndrome—highlights the way in which microbiome knowledge will enhance the precision, objectivity and fairness of our legal systems. Here in this opinion essay, we introduce the concept of the ‘legalome’—the application of microbiome and omics science to forensic psychiatry and criminal law. We argue that the rapid pace of microbial discoveries, including those that challenge ideas of free will and moral responsibility, will necessitate a reconsideration of traditional legal doctrines and justifications of retributive punishment. The implications extend beyond the courtroom, challenging us to reconsider how environmental factors—from diet to socioeconomic conditions—might shape preventative and rehabilitative efforts through their effects on the microbiome.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70129","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143595565","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}
Ivan Casas-Rodrigo, Tobias Vornholt, Kathrin Castiglione, Tania Michelle Roberts, Markus Jeschek, Thomas R. Ward, Sven Panke
{"title":"Permeabilisation of the Outer Membrane of Escherichia coli for Enhanced Transport of Complex Molecules","authors":"Ivan Casas-Rodrigo, Tobias Vornholt, Kathrin Castiglione, Tania Michelle Roberts, Markus Jeschek, Thomas R. Ward, Sven Panke","doi":"10.1111/1751-7915.70122","DOIUrl":"https://doi.org/10.1111/1751-7915.70122","url":null,"abstract":"<p>The bacterial envelope plays a critical role in maintaining essential cellular functions by selectively regulating import and export. The selectivity of this envelope can restrict the utilisation of externally provided compounds, thereby restricting the functional space of cellular engineering. This study systematically investigates the potential of large pore outer membrane proteins (OMPs) to enhance outer membrane permeability for diverse challenging compounds. We focus on the general porin OmpF, which facilitates the diffusion of water and small molecules, and specific OMP transporters FhuA and FepA, which mediate the translocation of small hydrophilic compounds. Through comprehensive characterisation, we evaluate the effects of recombinant expression of OMPs and engineered variants for small and hydrophilic compounds, aromatic molecules and bulky molecules and apply our findings to address two critical contemporary transport challenges: the uptake of large metal-containing cofactors for artificial metalloenzymes and non-permeant fluorescent Halo-ligands for in vivo protein labelling. Notably, we demonstrate significant improvements in ArM-catalysis and labelling. This study provides a practical guide for designing experiments that include outer-membrane-transport-limiting steps. This study highlights the potential of engineered OMPs to overcome the limitations imposed by the cell envelope, enabling the incorporation of complex molecules and expanding the frontiers of cellular engineering.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70122","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143581580","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}
Ivan Casas-Rodrigo, Tobias Vornholt, Kathrin Castiglione, Tania Michelle Roberts, Markus Jeschek, Thomas R. Ward, Sven Panke
{"title":"Permeabilisation of the Outer Membrane of Escherichia coli for Enhanced Transport of Complex Molecules","authors":"Ivan Casas-Rodrigo, Tobias Vornholt, Kathrin Castiglione, Tania Michelle Roberts, Markus Jeschek, Thomas R. Ward, Sven Panke","doi":"10.1111/1751-7915.70122","DOIUrl":"https://doi.org/10.1111/1751-7915.70122","url":null,"abstract":"<p>The bacterial envelope plays a critical role in maintaining essential cellular functions by selectively regulating import and export. The selectivity of this envelope can restrict the utilisation of externally provided compounds, thereby restricting the functional space of cellular engineering. This study systematically investigates the potential of large pore outer membrane proteins (OMPs) to enhance outer membrane permeability for diverse challenging compounds. We focus on the general porin OmpF, which facilitates the diffusion of water and small molecules, and specific OMP transporters FhuA and FepA, which mediate the translocation of small hydrophilic compounds. Through comprehensive characterisation, we evaluate the effects of recombinant expression of OMPs and engineered variants for small and hydrophilic compounds, aromatic molecules and bulky molecules and apply our findings to address two critical contemporary transport challenges: the uptake of large metal-containing cofactors for artificial metalloenzymes and non-permeant fluorescent Halo-ligands for in vivo protein labelling. Notably, we demonstrate significant improvements in ArM-catalysis and labelling. This study provides a practical guide for designing experiments that include outer-membrane-transport-limiting steps. This study highlights the potential of engineered OMPs to overcome the limitations imposed by the cell envelope, enabling the incorporation of complex molecules and expanding the frontiers of cellular engineering.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 3","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70122","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143581508","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}