Enzyme and Microbial Technology最新文献

{"title":"Isolation of an endophytic yeast for improving the antibacterial activity of water chestnut Jiaosu: Focus on variation of microbial communities.","authors":"Khadija Ei Galai, Wenna Dai, Cheng Qian, Jing Ye, Qin Zhang, Mengdie Gao, Xinyu Yang, Yanbin Li","doi":"10.1016/j.enzmictec.2025.110584","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2025.110584","url":null,"abstract":"<p><p>Recent years have seen an increase in the development of functional Jiaosu products, including eco-friendly Jiaosu and antimicrobial healthcare fermentation products. As a result, research on the antibacterial activity of Jiaosu has attracted attention. In the present study, the endophytic yeast WCF016, which exhibits antibacterial activity against Escherichia coli and Staphylococcus aureus, was isolated from the peel of water chestnut and identified as Candida sake via morphological and phylogenetic analyses based on 26S rDNA D1/D2 region sequencing. Water chestnut Jiaosu with or without WCF016 inoculation exhibited similar flavor and physicochemical properties. However, inoculation significantly enhanced the antibacterial activity of water chestnut Jiaosu, especially in group D (inoculate of both fruit and vegetable enzyme starter and WCF016), which showed the largest diameter in its inhibition zone for both E. coli and S. aureus, reaching 25 ± 0 mm and 24 ± 1.0 mm. Moreover, inoculation with WCF016 influenced the abundance of the microbial community, especially Lactiplantibacillus and Zygoascus, which reached 51.76 % and 24.46 %, respectively, in group B (inoculated WCF016), thereby improving the antibacterial activity and flavor quality of the water chestnut Jiaosu. Notably, final pH, total sugar, and all organic acids effectively promoted fungal diversity and exhibited a positive correlation with most of the fungal genera. These results indicate that conditions conducive to the formation of organic acid-producing microbes and the synthesis of organic acids promote the antibacterial activity of Jiaosu.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110584"},"PeriodicalIF":3.4,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cell-free biocatalysis for co-production of nicotinamide mononucleotide and ethanol from Saccharomyces cerevisiae and recombinant Escherichia coli.","authors":"Anoth Maharjan, Mamata Singhvi, Beom Soo Kim","doi":"10.1016/j.enzmictec.2025.110585","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2025.110585","url":null,"abstract":"<p><p>Cell-free enzyme systems have emerged as a promising approach for producing various biometabolites, offering several advantages over traditional whole-cell systems. This study presents an approach to producing nicotinamide mononucleotide (NMN) by combining a Saccharomyces cerevisiae cell-free enzyme with a recombinant Escherichia coli cell-free enzyme. The system leverages the ATP generated by yeast during ethanol fermentation to produce NMN in the presence of nicotinamide (NAM) as a substrate. The optimal cell-free enzyme concentration and substrate concentration were investigated to maximize NMN production. The results showed that combined cell-free enzymes led to increased NMN and ethanol yields, with a maximum production of 1.5 mM NMN (2.7-fold) and ethanol production of 0.45 g/L achieved (1.6-fold) compared to individual cell-free enzymes. Furthermore, the study demonstrated that the protein concentration affected NMN production, with optimal production achieved at 5 g/L. This study demonstrates the potential of integrating multiple metabolic pathways in a single cell-free system, paving the way for the development of more efficient and sustainable bioproduction processes.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110585"},"PeriodicalIF":3.4,"publicationDate":"2025-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of the support alkyl chain nature in the functional properties of the immobilized lipases.","authors":"Diandra de Andrades, Pedro Abellanas-Perez, Javier Rocha-Martin, Fernando Lopez-Gallego, Andrés R Alcántara, Maria de Lourdes Teixeira de Moraes Polizeli, Roberto Fernandez-Lafuente","doi":"10.1016/j.enzmictec.2025.110583","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2025.110583","url":null,"abstract":"<p><p>Supports coated with amino-hexyl and amino octyl have been prepared from glyoxyl agarose beads and compared in their performance with octyl-agarose to immobilize lipases A and B from Candida antarctica (CALA and CALB). Immobilization courses were similar using all supports, but enzyme release was more difficult using the amino-alkyl supports suggesting a mixed interfacial activation/ionic exchange immobilization. The enzyme activity and specificity (using p-nitrophenyl propionate, triacetin and both isomers of methyl mandelate) greatly depended on the support. In many instances the enzymes immobilized on the new supports offered higher activities and enantiospecificity in the hydrolysis of both enantiomers of methyl mandelate (mainly using CALB). This was coupled to a lower enzyme stability using the new supports, even in the presence of high ionic strength, suggesting that the amphipathic could be responsible of the enzyme lower stability. Using CALB, it was possible to detect a higher exposition of the enzyme Trp groups to the medium by florescence spectra after its immobilization on the amino-alkyl-supports, correlating to the higher activity and lower stability results.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110583"},"PeriodicalIF":3.4,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biosynthesis of 2-phenylethanol from styrene using engineered Escherichia coli whole cells.","authors":"Tianzhen Xiong, Qiuyue Gao, Wei Liu, Wei Li, Guangyan Fan","doi":"10.1016/j.enzmictec.2025.110582","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2025.110582","url":null,"abstract":"<p><p>2-Phenylethanol, an aromatic alcohol with a rose scent, is widely used in the cosmetics, food, and pharmaceutical industries. We designed an efficient multi-enzyme cascade pathway for production of 2-phenylethanol from styrene as the substrate. Initially, 2-phenylethanol was produced by overexpression of styrene monooxygenase A (styA), styrene monooxygenase B (styB), styrene oxide isomerase (SOI), alcohol dehydrogenase (yahK), and glucose dehydrogenase (gdh) in Escherichia coli to give 6.28 mM 2-phenylethanol. Subsequently, plasmids with different copy numbers were employed to balance the expression of pathway enzymes to produce 10.28 mM 2-phenylethanol, resulting in a 63.7 % increase in the final yield. Furthermore, the pH and temperature of the whole-cell conversion reaction were optimized, the optimum pH and temperature are 7.5 and 35℃, respectively. Finally, whole-cell conversion experiment was conducted, and the production of 2-phenylethanol reached 48.17 mM within 10 h. This study provides a theoretical and practical foundation for production of 2-phenylethanol.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110582"},"PeriodicalIF":3.4,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142970129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of enzyme kinetics of fungal methionine synthases in an optimized colorimetric microscale assay for measuring cobalamin-independent methionine synthase activity.","authors":"Noël Jung, Tomás Vellozo-Echevarría, Kristian Barrett, Anne S Meyer","doi":"10.1016/j.enzmictec.2025.110581","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2025.110581","url":null,"abstract":"<p><p>Aspergillus spp. and Rhizopus spp., used in solid-state plant food fermentations, encode cobalamin-independent methionine synthase activity (MetE, EC 2.1.1.14). Here, we examine the enzyme kinetics, reaction activation energies (E_a), thermal robustness, and structural folds of three MetEs from three different food-fermentation relevant fungi, Aspergillus sojae, Rhizopus delemar, and Rhizopus microsporus, and compare them to the MetE from Escherichia coli. We also downscaled and optimized a colorimetric assay to allow direct MetE activity measurements in microplates. The catalytic rates, k_cat, of the three fungal MetE enzymes on the methyl donor (6S)-5-methyl-tetrahydropteroyl-L-glutamate₃ ranged from 1.2 to 3.3 min^-1 and K_M values varied from 0.8 to 6.8 µM. The k_cat was lowest for the R. delemar MetE, but this enzyme also had the lowest K_M thus resulting in the highest k_cat/K_M of ∼1.4 min^-1 µM^-1 among the three fungal enzymes. The k_cat was higher for the E. coli enzyme, 12 min^-1, but K_M was 6.4 µM, resulting in k_cat/K_M of ∼1.9 min^-1 µM^-1. The E_a values of the fungal MetEs ranged from 52 to 97 kJ mole^-1 and were higher than that of the E. coli MetE (38.7 kJ mole ^-1). The predicted structural folds of the MetEs were very similar. T_m values of the fungal MetEs ranged from 41 to 54 °C, highest for the A. sojae enzyme (54 °C), lowest for the R. delemar (41 °C). At 30 °C, the half-lives of the three fungal enzymes varied significantly, with MetE from A. sojae having the longest (> 600 min, k_D=0), and R. delemar the shortest (17 min). Knowledge of the kinetics of these enzymes is important for understanding methionine synthesis in fungi and a first step in promoting methionine synthesis in fungally fermented plant foods.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110581"},"PeriodicalIF":3.4,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering glycolytic pathway for improved Lacto-N-neotetraose production in pichia pastoris.","authors":"Jiao Yang, Nitesh Kumar Mund, Lirong Yang, Hao Fang","doi":"10.1016/j.enzmictec.2024.110576","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110576","url":null,"abstract":"<p><p>Lacto-N-neotetraose (LNnT) is a primary solid component of human milk oligosaccharides (HMOs) with various promising health effects for infants. LNnT production by GRAS (generally recognized as safe) microorganisms has attracted considerable attention. However, few studies have emphasized Pichia Pastoris as a cell factory for LNnT's production. Here, we have reported the first-ever synthesis of LNnT employing P. pastoris as the host. Initially, LNnT biosynthetic pathway genes β-1,3-N-acetylglucosaminyltransferase (lgtA) and β-1,4-galactostltransferase (lgtB) along with lactose permease (lac12) and galactose epimerase (gal10) were integrated into the genome of P. pastoris, but only 0.139 g/L LNnT was obtained. Second, the titer of LNnT was improved to 0.162 g/L via up-regulating genes to strengthen the supply of precursors, UDP-GlcNAc (Uridine diphosphate N-acetylglucosamine) and UDP-Gal (Uridine diphosphate galactose), for LNnT biosynthesis. Third, by knocking out critical mediator pfk (6-phosphofructokinase) genes in glycolysis, the major glucose metabolic flux was rewired to the LNnT biosynthesis pathway. As a result, the strain accumulated 0.867 g/L LNnT in YPG medium supplemented with glucose and lactose. Finally, LNnT production was increased to 1.24 g/L in a 3 L bioreactor. The work aimed to explore the potential of P. pastoris as a for LNnT production.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110576"},"PeriodicalIF":3.4,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142913974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structures and properties of α-amylase and glucoamylase immobilized by ZIF-8 via one-pot preparation.","authors":"Yuxin Liu, Qinghua Pan, Zesheng Liang, Jingqiao Li, Rulong Wu","doi":"10.1016/j.enzmictec.2024.110579","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110579","url":null,"abstract":"<p><p>The immobilization of α-amylase and glucoamylase using a metal-organic framework (enzyme@ZIF-8) was prepared in situ through a one-pot method. The morphology, crystal structure, and molecular characteristics of the free enzyme and enzyme@ZIF-8 were characterized. The enzyme@ZIF-8 exhibited the rhombic dodecahedron morphology, with a decrease in particle size. Successful immobilization of α-amylase and glucoamylase within ZIF-8 was confirmed, with 30-40 % loading rate. The immobilization process did not significantly alter the crystal structure of ZIF-8. The changes in secondary structure of enzyme after immobilization resulted in modification of catalytic activity of enzyme. The melting enthalpy of enzyme @ZIF-8 increased with the increase of enzyme content. The melting peak temperature of the enzyme immobilized by ZIF-8 increased. The activity of free and immobilized enzymes was influenced by the different time, pH and temperature. At pH 5-8 and temperature 60-80 °C, the activity of the immobilized enzyme was significantly greater than that of the free enzyme. The repeatability of enzyme@ZIF-8 was 61.52 % after three cycles. The kinetic parameters of Michaelis-Menten model for enzymatic reaction were determined by fitting the initial rate of reactions and initial substrate concentration data. The Michaelis-Menten constant (K_M) values of immobilized enzyme were lower than that of free enzyme, indicating the greater affinity between the enzyme and the substrate.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110579"},"PeriodicalIF":3.4,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142926836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of a bacteria P450 enzyme from B. megaterium H-1 with vitamin D₃ C-25 hydroxylation capabilities.","authors":"Yulin He, Yina Hou, Hui Li, Fan He, Jingyi Zhou, Xiaomei Zhang, Jingsong Shi, Zhenghong Xu","doi":"10.1016/j.enzmictec.2024.110578","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110578","url":null,"abstract":"<p><p>Calcidiol (25(OH)VD₃) and calcitriol (1α,25(OH)₂VD₃) are active vitamin D₃ with high medicinal value, which can maintain calcium and phosphorus balance and treat vitamin D deficiency. Microbial synthesis is an important method to produce high-value-added compounds. It can produce active vitamin D₃ through the hydroxylation reaction of P450, which can reduce the traditional chemical synthesis steps, and greatly improve the production efficiency and economic benefits. In this work, Bacillus megaterium H-1 was screened for its ability to produce 25(OH)VD₃ and 1α,25(OH)₂VD₃ from vitamin D₃. A new highly inducible vitamin D₃ hydroxylase CYP109E1-H was identified from B. megaterium H-1 through searching for transcripts with cytochrome P450 structural domains, combining the transcriptome sequencing with functional expression in Bacillus subtilis WB600. Biotransformation in recombinant B. subtilis confirmed that CYP109E1-H has C-25 hydroxylase activity towards vitamin D₃. CYP109E1-H is a natural mutant of CYP109E1 with greater stereoselectivity and it is a new vitamin D₃ mono-hydroxylase. The cloning and characterization of the CYP109E1-H gene provide useful information on the structural basis for improving the regional and stereoselectivity of the CYP109E gene.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110578"},"PeriodicalIF":3.4,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142892980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acidogenic fermentation of Ulva in a fed-batch reactor system: tubular versus foliose biomass.","authors":"James Lawrence, Armando Oliva, Jerry D Murphy, Piet N L Lens","doi":"10.1016/j.enzmictec.2024.110580","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110580","url":null,"abstract":"<p><p>The present study proposes a biorefinery of the macroalgae Ulva, focusing on evaluating two different morphologies of the species (foliose and tubular) during acidogenic fermentation in fed-batch reactors. Stage 1 of the study evaluates lyophilised foliose and tubular Ulva, whilst Stage 2 analyses the impact of ulvan extraction on volatile fatty acids yield and changes in carbohydrate availability. Acetic, propionic, and butyric acids were produced from each substrate, with peak concentrations of total VFAs recorded at 2179.5 mg HAc/L (foliose Ulva) and 2029.3 mg HAc/L (tubular Ulva) when ulvan was present. After ulvan extraction, the acidogenic fermentation of the foliose morphotype was negatively affected, reaching at most 315.3 mg HAc/L. In contrast, the extraction showed no influence on the tubular morphotype, peaking at 2165.0 mg HAc/L. Additional variations were noted in the availability of carbohydrates in each substrate during the acidogenic fermentation process. The ulvan-extracted tubular morphotype exhibited the highest peak in carbohydrate concentration (9.8 g glucose/L), whilst the ulvan-extracted foliose morphotype yielded up to 8.5 g glucose/L. This study highlights the biorefinery potential of Ulva biomass, proposing a multiple cascading approach linking multiple energy and biomolecule applications to maximise the valorisation of the biomass.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110580"},"PeriodicalIF":3.4,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142946515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Green and sustainable production of isofraxidin from Acanthopanax senticosus with cellulose-based immobilized probiotics.","authors":"Shuang Jin, Hongyao Cai, Cailiang Peng, Yupeng Cheng, Yubin Ren, Weili Liu, Yujie Fu, Chen Lv","doi":"10.1016/j.enzmictec.2024.110574","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110574","url":null,"abstract":"<p><p>This study utilizes deep eutectic solvent (DES)-assisted enhancement of cellulose-based immobilized probiotics for efficient biotransformation of isofraxidin from Acanthopanax senticosus. Among seven probiotic strains tested, Lactiplantibacillus plantarum CICC 20767 exhibited the best catalytic activity. We explored the effects of 12 different DESs with varying hydrogen bond donors on the conversion capacity of the immobilized probiotics, with the highest efficiency observed using ChCl/EG (4.0 wt %). The optimized process, with a solid-to-liquid ratio of 1:5 (g/mL), a temperature of 35.6 °C, a reaction time of 4 d, and a pH of 6.9, resulted in a 5.53-folds increase in isofraxidin yield, reaching 0.4034 mg/g, compared to the untreated sample (0.0729 mg/g). The immobilized probiotics retained excellent catalytic activity after 12 cycles of use, demonstrating their stability and potential for large-scale, green production of isofraxidin. This study presents a valuable method for industrial isofraxidin production and highlights the broad potential of this environmentally friendly bioconversion process in the pharmaceutical industry.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110574"},"PeriodicalIF":3.4,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142892979","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}