Pub Date : 2024-05-22DOI: 10.3390/fermentation10060270
Marina Grubišić, Ines Peremin, Elvis Djedović, B. Šantek, Mirela Ivančić Šantek
Microalgal biomass is an excellent platform for producing food, feed, nutraceuticals, pharmaceuticals, and biofuels. This study aimed to investigate the effect of the trophic mode of cultivation (phototrophic, heterotrophic, and mixotrophic) on the growth and biomass composition of Chlorella vulgaris S2. The contents of lipids and carbohydrates, as well as the fatty acid composition of total lipids, were studied. The effects of the carbon-to-nitrogen ratio (C:N) and the organic carbon concentration of the growth media under mixotrophic and heterotrophic conditions were also investigated. The C:N ratio of 30 mol mol−1 favoured lipid synthesis, and the C:N ratio of 10 mol mol−1 favoured carbohydrate synthesis. Maximal lipid and biomass productivities (2.238 and 0.458 g L−1 d−1, respectively) were obtained under mixotrophic conditions at the C:N ratio of 50 mol mol−1 and glucose concentration of 50 g L−1. Fed-batch cultivation conducted in a stirrer tank bioreactor under heterotrophic growth conditions increased biomass (2.385 g L−1 d−1, respectively) and lipid (0.339 L−1 d−1) productivities ~50 and ~60 times compared to the fed-batch phototrophic cultivation, respectively. The trophic mode, growth phase, and growth medium composition significantly influenced the fatty acid composition. Under mixotrophic and heterotrophic growth conditions, lipid accumulation is associated with an increase in oleic acid (C18:1) content. Mixotrophically grown biomass of Chlorella vulgaris S2 under optimised conditions is a suitable source of lipids for biodiesel production.
{"title":"Cultivation of a Novel Strain of Chlorella vulgaris S2 under Phototrophic, Mixotrophic, and Heterotrophic Conditions, and Effects on Biomass Growth and Composition","authors":"Marina Grubišić, Ines Peremin, Elvis Djedović, B. Šantek, Mirela Ivančić Šantek","doi":"10.3390/fermentation10060270","DOIUrl":"https://doi.org/10.3390/fermentation10060270","url":null,"abstract":"Microalgal biomass is an excellent platform for producing food, feed, nutraceuticals, pharmaceuticals, and biofuels. This study aimed to investigate the effect of the trophic mode of cultivation (phototrophic, heterotrophic, and mixotrophic) on the growth and biomass composition of Chlorella vulgaris S2. The contents of lipids and carbohydrates, as well as the fatty acid composition of total lipids, were studied. The effects of the carbon-to-nitrogen ratio (C:N) and the organic carbon concentration of the growth media under mixotrophic and heterotrophic conditions were also investigated. The C:N ratio of 30 mol mol−1 favoured lipid synthesis, and the C:N ratio of 10 mol mol−1 favoured carbohydrate synthesis. Maximal lipid and biomass productivities (2.238 and 0.458 g L−1 d−1, respectively) were obtained under mixotrophic conditions at the C:N ratio of 50 mol mol−1 and glucose concentration of 50 g L−1. Fed-batch cultivation conducted in a stirrer tank bioreactor under heterotrophic growth conditions increased biomass (2.385 g L−1 d−1, respectively) and lipid (0.339 L−1 d−1) productivities ~50 and ~60 times compared to the fed-batch phototrophic cultivation, respectively. The trophic mode, growth phase, and growth medium composition significantly influenced the fatty acid composition. Under mixotrophic and heterotrophic growth conditions, lipid accumulation is associated with an increase in oleic acid (C18:1) content. Mixotrophically grown biomass of Chlorella vulgaris S2 under optimised conditions is a suitable source of lipids for biodiesel production.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"27 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141110904","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-21DOI: 10.3390/fermentation10060267
Jourdan E. Lakes, Brittany E. Davis, M. Flythe
Amino-acid-fermenting bacteria are wasteful organisms within the rumens of beef cattle that remove dietary amino nitrogen by producing ammonia, which is then excreted renally. There are currently no on-label uses for the control of this microbial guild, but off-label use of broad-spectrum antimicrobials has shown efficacy, which contributes to antimicrobial resistance. Plant-derived antimicrobials supplemented into the diets of cattle may offer worthwhile alternatives. This study sought to investigate the role of cannabidiol (CBD) as a terpenophenolic antimicrobial. Ex vivo cell suspensions were harvested from the rumen fluid of Angus × Holstein steers in non-selective media with amino acid substrates. The suspensions were treated with five concentrations of CBD (860 μg mL−1–0.086 μg mL−1) and incubated (24 h), after which ammonia production and viable number of cells per substrate and treatment were measured. The data demonstrated a ~10–15 mM reduction in ammonia produced at the highest concentration of CBD and negligible changes in the viable number of amino-acid-fermenting bacteria. CBD does not appear to be a biologically or economically viable terpenophenolic candidate for the control of amino acid fermentation in beef cattle.
{"title":"Activity of Cannabidiol on Ex Vivo Amino Acid Fermentation by Bovine Rumen Microbiota","authors":"Jourdan E. Lakes, Brittany E. Davis, M. Flythe","doi":"10.3390/fermentation10060267","DOIUrl":"https://doi.org/10.3390/fermentation10060267","url":null,"abstract":"Amino-acid-fermenting bacteria are wasteful organisms within the rumens of beef cattle that remove dietary amino nitrogen by producing ammonia, which is then excreted renally. There are currently no on-label uses for the control of this microbial guild, but off-label use of broad-spectrum antimicrobials has shown efficacy, which contributes to antimicrobial resistance. Plant-derived antimicrobials supplemented into the diets of cattle may offer worthwhile alternatives. This study sought to investigate the role of cannabidiol (CBD) as a terpenophenolic antimicrobial. Ex vivo cell suspensions were harvested from the rumen fluid of Angus × Holstein steers in non-selective media with amino acid substrates. The suspensions were treated with five concentrations of CBD (860 μg mL−1–0.086 μg mL−1) and incubated (24 h), after which ammonia production and viable number of cells per substrate and treatment were measured. The data demonstrated a ~10–15 mM reduction in ammonia produced at the highest concentration of CBD and negligible changes in the viable number of amino-acid-fermenting bacteria. CBD does not appear to be a biologically or economically viable terpenophenolic candidate for the control of amino acid fermentation in beef cattle.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"134 46","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141115185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To enhance comprehension of the impact of mixed fermentation using Torulaspora delbrueckii Bio-119667 (TD) on the aroma composition of Munage grape base-wine (MGBW), we analysed the aroma composition of MGBW using HS-SPME-GC-MS widely targeted metabolomics. The levels of volatile aroma components, including terpenes, higher alcohols, aldehydes, heterocyclic compounds, and esters, were significantly higher in MGBW produced by mixed fermentation compared to the pure Saccharomyces yeast control fermentation. The study found that the content of esters increased by 26.3% after mixed fermentation, indicating the contribution of TD to the formation of ester flavour components during the fermentation of MGBW. After analysing aroma activity values, we discovered that 49 out of 115 esters (25.5% of the total) significantly contributed to the aroma profile of MGBW (rOAV > 1). Of these esters, 16 were identified as key aroma compounds (rOAV > 1, VIP > 1) produced by mixed fermentation with the participation of TD. This finding further supports the contribution of TD to the improvement of MGBW’s aroma composition. This study reveals the role of non-Saccharomyces yeast strain Torulaspora delbrueckii Bio-119667 in improving the aroma composition of MGBW produced by mixed culture fermentation and the biosynthetic pathways of key aroma components therein.
{"title":"The Effect of Non-Saccharomyces Cerevisiae Torulaspora delbrueckii on the Aroma Composition of Munage Grape Base-Wine and the Mechanism of the Effect","authors":"Xuefeng Yin, Bingze Liu, Ruxianguli Maimaitiyiming, Liang Wang, Lei Zhao, Huimin Zhang, Keping Chen, Aihemaitijiang Aihaiti","doi":"10.3390/fermentation10050266","DOIUrl":"https://doi.org/10.3390/fermentation10050266","url":null,"abstract":"To enhance comprehension of the impact of mixed fermentation using Torulaspora delbrueckii Bio-119667 (TD) on the aroma composition of Munage grape base-wine (MGBW), we analysed the aroma composition of MGBW using HS-SPME-GC-MS widely targeted metabolomics. The levels of volatile aroma components, including terpenes, higher alcohols, aldehydes, heterocyclic compounds, and esters, were significantly higher in MGBW produced by mixed fermentation compared to the pure Saccharomyces yeast control fermentation. The study found that the content of esters increased by 26.3% after mixed fermentation, indicating the contribution of TD to the formation of ester flavour components during the fermentation of MGBW. After analysing aroma activity values, we discovered that 49 out of 115 esters (25.5% of the total) significantly contributed to the aroma profile of MGBW (rOAV > 1). Of these esters, 16 were identified as key aroma compounds (rOAV > 1, VIP > 1) produced by mixed fermentation with the participation of TD. This finding further supports the contribution of TD to the improvement of MGBW’s aroma composition. This study reveals the role of non-Saccharomyces yeast strain Torulaspora delbrueckii Bio-119667 in improving the aroma composition of MGBW produced by mixed culture fermentation and the biosynthetic pathways of key aroma components therein.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"91 22","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141122953","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-19DOI: 10.3390/fermentation10050264
N. Kouadio, Alalet Luc Olivier Zady, Kouassi Aboutou Sévérin Kra, Filofteia Camelia Diguță, S. Niamké, F. Matei
This study aimed to evaluate the probiotic properties of 10 lactic acid bacteria (LAB) isolated from artisanal fermented plantain dockounou paste. A preliminary characterization of the LAB isolates was performed based on phenotypic and several biochemical properties, which was subsequently confirmed through 16S rRNA gene sequencing analysis, indicating that these isolates belonged to the species Lactiplantibacillus plantarum. With regard to safety criteria, the strains exhibited no alpha or beta hemolysis activity. Nevertheless, the majority of LAB strains demonstrated high sensitivity to the antibiotics tested. The results demonstrated that the majority of the strains exhibited remarkably high survival rates under simulated gastrointestinal conditions, such as pH = 1.5 (81.18–98.15%), 0.3% bile salts (68.62–100.89%), 0.4% phenol (40.59–128.24%), as well as 0.1% pepsin and pH = 2.5 (88.54–99.78%). The LAB strains demonstrated elevated levels of cell surface properties, indicative of the presence of a considerable defensive mechanism against pathogens. Intact LAB cells exhibited significant antioxidant abilities (48.18–83.58%). They also demonstrated a pronounced inhibitory effect on the growth of foodborne pathogens. Enzyme pattern analysis revealed that the LAB isolates produced both proteases and cellulases, as well as pectinase and/or amylase activity. The potential of the L. plantarum strains FS43, FS44, and FS48, as indicated by the results obtained from the standard in vitro assays, makes them suitable for further study as potential probiotics.
{"title":"In Vitro Probiotic Characterization of Lactiplantibacillus plantarum Strains Isolated from Traditional Fermented Dockounou Paste","authors":"N. Kouadio, Alalet Luc Olivier Zady, Kouassi Aboutou Sévérin Kra, Filofteia Camelia Diguță, S. Niamké, F. Matei","doi":"10.3390/fermentation10050264","DOIUrl":"https://doi.org/10.3390/fermentation10050264","url":null,"abstract":"This study aimed to evaluate the probiotic properties of 10 lactic acid bacteria (LAB) isolated from artisanal fermented plantain dockounou paste. A preliminary characterization of the LAB isolates was performed based on phenotypic and several biochemical properties, which was subsequently confirmed through 16S rRNA gene sequencing analysis, indicating that these isolates belonged to the species Lactiplantibacillus plantarum. With regard to safety criteria, the strains exhibited no alpha or beta hemolysis activity. Nevertheless, the majority of LAB strains demonstrated high sensitivity to the antibiotics tested. The results demonstrated that the majority of the strains exhibited remarkably high survival rates under simulated gastrointestinal conditions, such as pH = 1.5 (81.18–98.15%), 0.3% bile salts (68.62–100.89%), 0.4% phenol (40.59–128.24%), as well as 0.1% pepsin and pH = 2.5 (88.54–99.78%). The LAB strains demonstrated elevated levels of cell surface properties, indicative of the presence of a considerable defensive mechanism against pathogens. Intact LAB cells exhibited significant antioxidant abilities (48.18–83.58%). They also demonstrated a pronounced inhibitory effect on the growth of foodborne pathogens. Enzyme pattern analysis revealed that the LAB isolates produced both proteases and cellulases, as well as pectinase and/or amylase activity. The potential of the L. plantarum strains FS43, FS44, and FS48, as indicated by the results obtained from the standard in vitro assays, makes them suitable for further study as potential probiotics.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"110 25","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141124660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-19DOI: 10.3390/fermentation10050265
S. But, R. Z. Suleimanov, I. Y. Oshkin, O. Rozova, I. Mustakhimov, Nikolai V. Pimenov, S. Dedysh, V. Khmelenina
The biotechnology of converting methane to single-cell protein (SCP) implies using fast-growing thermotolerant aerobic methanotrophic bacteria. Among the latter, members of the genus Methylococcus received significant research attention and are used in operating commercial plants. Methylococcus capsulatus MIR is a recently discovered member of this genus with the potential to be used for the purpose of SCP production. Like other Methylococcus species, this bacterium stores carbon and energy in the form of glycogen, particularly when grown under nitrogen-limiting conditions. The genome of strain MIR encodes two glycogen synthases, GlgA1 and GlgA2, which are only moderately related to each other. To obtain glycogen-free cell biomass of this methanotroph, glycogen synthase mutants, ΔglgA1, ΔglgA2, and ΔglgA1ΔglgA2, were constructed. The mutant lacking both glycogen synthases exhibited a glycogen-deficient phenotype, whereas the intracellular glycogen content was not reduced in strains defective in either GlgA1 or GlgA2, thus suggesting functional redundancy of these enzymes. Inactivation of the glk gene encoding glucokinase also resulted in a sharp decrease in glycogen content and accumulation of free glucose in cells. Wild-type strain MIR and the mutant strain ΔglgA1ΔglgA2 were also grown in a bioreactor operated in batch and continuous modes. Cell biomass of ΔglgA1ΔglgA2 mutant obtained during batch cultivation displayed high protein content (71% of dry cell weight (DCW) compared to 54% DCW in wild-type strain) as well as a strong reduction in glycogen content (10.8 mg/g DCW compared to 187.5 mg/g DCW in wild-type strain). The difference in protein and glycogen contents in biomass of these strains produced during continuous cultivation was less pronounced, yet biomass characteristics relevant to SCP production were slightly better for ΔglgA1ΔglgA2 mutant. Genome analysis revealed the presence of glgA1-like genes in all methanotrophs of the Gammaproteobacteria and Verrucomicrobia, while only a very few methanotrophic representatives of the Alphaproteobacteria possessed these determinants of glycogen biosynthesis. The glgA2-like genes were present only in genomes of gammaproteobacterial methanotrophs with predominantly halo- and thermotolerant phenotypes. The role of glycogen in terms of energy reserve is discussed.
将甲烷转化为单细胞蛋白（SCP）的生物技术意味着要使用快速生长的耐高温好氧甲烷营养细菌。其中，甲基球菌属（Methylococcus）的成员受到了大量研究的关注，并被用于商业工厂的运营。荚膜甲基球菌 MIR 是最近发现的该属的一个成员，有可能用于生产 SCP。与其他甲基球菌一样，这种细菌以糖原的形式储存碳和能量，尤其是在氮限制条件下生长时。菌株 MIR 的基因组编码两种糖原合成酶，即 GlgA1 和 GlgA2，它们之间的亲缘关系一般。为了获得这种甲烷营养体的无糖原细胞生物量，构建了糖原合成酶突变体，即ΔglgA1、ΔglgA2和ΔglgA1ΔglgA2。缺乏这两种糖原合成酶的突变体表现出糖原缺乏的表型，而 GlgA1 或 GlgA2 均有缺陷的菌株细胞内糖原含量并不减少，因此表明这些酶存在功能冗余。编码葡萄糖激酶的 glk 基因失活也会导致糖原含量急剧下降和细胞内游离葡萄糖的积累。野生型菌株 MIR 和突变型菌株 ΔglgA1ΔglgA2 也在以间歇和连续模式运行的生物反应器中生长。在批量培养过程中获得的突变株ΔglgA1ΔglgA2的细胞生物量显示出较高的蛋白质含量（占细胞干重（DCW）的71%，而野生型菌株的DCW为54%），以及糖原含量的显著降低（10.8 mg/g DCW，而野生型菌株的DCW为187.5 mg/g）。这些菌株在连续培养过程中产生的生物量中蛋白质和糖原含量的差异不太明显，但ΔglgA1ΔglgA2突变体与SCP生产相关的生物量特征略好。基因组分析表明，在所有产甲烷的加玛蛋白菌和Verrucomicrobia中都存在类似glgA1的基因，而只有极少数产甲烷的阿尔法蛋白菌具有这些糖原生物合成的决定性基因。类似 glgA2 的基因只存在于主要具有耐晕和耐热表型的甘蛋白细菌甲烷营养体的基因组中。讨论了糖原在能量储备方面的作用。
{"title":"New Solutions in Single-Cell Protein Production from Methane: Construction of Glycogen-Deficient Mutants of Methylococcus capsulatus MIR","authors":"S. But, R. Z. Suleimanov, I. Y. Oshkin, O. Rozova, I. Mustakhimov, Nikolai V. Pimenov, S. Dedysh, V. Khmelenina","doi":"10.3390/fermentation10050265","DOIUrl":"https://doi.org/10.3390/fermentation10050265","url":null,"abstract":"The biotechnology of converting methane to single-cell protein (SCP) implies using fast-growing thermotolerant aerobic methanotrophic bacteria. Among the latter, members of the genus Methylococcus received significant research attention and are used in operating commercial plants. Methylococcus capsulatus MIR is a recently discovered member of this genus with the potential to be used for the purpose of SCP production. Like other Methylococcus species, this bacterium stores carbon and energy in the form of glycogen, particularly when grown under nitrogen-limiting conditions. The genome of strain MIR encodes two glycogen synthases, GlgA1 and GlgA2, which are only moderately related to each other. To obtain glycogen-free cell biomass of this methanotroph, glycogen synthase mutants, ΔglgA1, ΔglgA2, and ΔglgA1ΔglgA2, were constructed. The mutant lacking both glycogen synthases exhibited a glycogen-deficient phenotype, whereas the intracellular glycogen content was not reduced in strains defective in either GlgA1 or GlgA2, thus suggesting functional redundancy of these enzymes. Inactivation of the glk gene encoding glucokinase also resulted in a sharp decrease in glycogen content and accumulation of free glucose in cells. Wild-type strain MIR and the mutant strain ΔglgA1ΔglgA2 were also grown in a bioreactor operated in batch and continuous modes. Cell biomass of ΔglgA1ΔglgA2 mutant obtained during batch cultivation displayed high protein content (71% of dry cell weight (DCW) compared to 54% DCW in wild-type strain) as well as a strong reduction in glycogen content (10.8 mg/g DCW compared to 187.5 mg/g DCW in wild-type strain). The difference in protein and glycogen contents in biomass of these strains produced during continuous cultivation was less pronounced, yet biomass characteristics relevant to SCP production were slightly better for ΔglgA1ΔglgA2 mutant. Genome analysis revealed the presence of glgA1-like genes in all methanotrophs of the Gammaproteobacteria and Verrucomicrobia, while only a very few methanotrophic representatives of the Alphaproteobacteria possessed these determinants of glycogen biosynthesis. The glgA2-like genes were present only in genomes of gammaproteobacterial methanotrophs with predominantly halo- and thermotolerant phenotypes. The role of glycogen in terms of energy reserve is discussed.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141123687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-18DOI: 10.3390/fermentation10050263
B. Zieniuk, Karina Jasińska, K. Wierzchowska, Şuheda Uğur, A. Fabiszewska
Yarrowia lipolytica is a robust yeast species that has gained significant attention as a biofactory for various biotechnological applications and undoubtedly can be referred to as a hidden treasure trove due to boasting a diverse array of enzymes with wide-ranging applications in multiple industries, including biofuel production, food processing, biotechnology, and pharmaceuticals. As the biotechnology field continues to expand, Y. lipolytica is poised to play a pivotal role in developing eco-friendly and economically viable bioprocesses. Its versatility and potential for large-scale production make it a promising candidate for sustainably addressing various societal and industrial needs. The current review article aimed to highlight the diverse enzymatic capabilities of Y. lipolytica and provide a detailed analysis of its relevance in biocatalysis, including the use of whole-cell catalysts and isolated enzymes. The review focused on wild-type yeast strains and their species-dependant properties and selected relevant examples of Y. lipolytica used as a host organism for overexpressing some enzymes. Furthermore, the application of Y. lipolytica’s potential in enantiomers resolution, lipids processing, and biodiesel synthesis, as well as the synthesis of polymers or esterification of different substrates for upgrading biologically active compounds, was discussed.
{"title":"Yarrowia lipolytica Yeast: A Treasure Trove of Enzymes for Biocatalytic Applications—A Review","authors":"B. Zieniuk, Karina Jasińska, K. Wierzchowska, Şuheda Uğur, A. Fabiszewska","doi":"10.3390/fermentation10050263","DOIUrl":"https://doi.org/10.3390/fermentation10050263","url":null,"abstract":"Yarrowia lipolytica is a robust yeast species that has gained significant attention as a biofactory for various biotechnological applications and undoubtedly can be referred to as a hidden treasure trove due to boasting a diverse array of enzymes with wide-ranging applications in multiple industries, including biofuel production, food processing, biotechnology, and pharmaceuticals. As the biotechnology field continues to expand, Y. lipolytica is poised to play a pivotal role in developing eco-friendly and economically viable bioprocesses. Its versatility and potential for large-scale production make it a promising candidate for sustainably addressing various societal and industrial needs. The current review article aimed to highlight the diverse enzymatic capabilities of Y. lipolytica and provide a detailed analysis of its relevance in biocatalysis, including the use of whole-cell catalysts and isolated enzymes. The review focused on wild-type yeast strains and their species-dependant properties and selected relevant examples of Y. lipolytica used as a host organism for overexpressing some enzymes. Furthermore, the application of Y. lipolytica’s potential in enantiomers resolution, lipids processing, and biodiesel synthesis, as well as the synthesis of polymers or esterification of different substrates for upgrading biologically active compounds, was discussed.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"104 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141124894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-04-25DOI: 10.3390/fermentation10050229
Nan Li, Lu Yang, Xiankun Ren, Peng Du, Piwu Li, Jing Su, Jing Xiao, Junqing Wang, Ruiming Wang
To explore the key sites affecting the intracellular assembly of key components of cellulosomes and obtain DocA mutants independent of Ca2+, Swiss-model, GROMACS, PyMOL, and other molecular dynamics simulation software were used for modeling and static and dynamic combination analysis. Site-specific mutation technology was used to mutate DocA, and Biacore was used to test the dependence of Ca2+ on the binding ability of protein DocA mutants and protein Coh, and to analyze the interaction and binding effect of mutant proteins in vitro. Forward intracellular mutant screening was performed based on semi-rational design and high throughput screening techniques. The orientation of mutations suitable for intracellular assembly was determined, and three directional mutant proteins, DocA-S1, DocA-S2, and DocA-S3, were obtained. Ca2+ independent DocA mutants were obtained gradually and their potential interaction mechanisms were analyzed. In the present study, intracellular self-assembly of key components of cellulosomes independent of Ca2+ was achieved, and DocA-S3 was applied to the assembly of key enzymes of L-lysine biosynthesis, in which DapA and DapB intracellular assembly increased L-lysine accumulation by 29.8% when compared with the control strains, providing a new strategy for improving the intracellular self-assembly of cellulosomes and amino acid fermentation efficiency.
{"title":"Improving the Synthesis Efficiency of Amino Acids by Analyzing the Key Sites of Intracellular Self-Assembly of Artificial Cellulosome","authors":"Nan Li, Lu Yang, Xiankun Ren, Peng Du, Piwu Li, Jing Su, Jing Xiao, Junqing Wang, Ruiming Wang","doi":"10.3390/fermentation10050229","DOIUrl":"https://doi.org/10.3390/fermentation10050229","url":null,"abstract":"To explore the key sites affecting the intracellular assembly of key components of cellulosomes and obtain DocA mutants independent of Ca2+, Swiss-model, GROMACS, PyMOL, and other molecular dynamics simulation software were used for modeling and static and dynamic combination analysis. Site-specific mutation technology was used to mutate DocA, and Biacore was used to test the dependence of Ca2+ on the binding ability of protein DocA mutants and protein Coh, and to analyze the interaction and binding effect of mutant proteins in vitro. Forward intracellular mutant screening was performed based on semi-rational design and high throughput screening techniques. The orientation of mutations suitable for intracellular assembly was determined, and three directional mutant proteins, DocA-S1, DocA-S2, and DocA-S3, were obtained. Ca2+ independent DocA mutants were obtained gradually and their potential interaction mechanisms were analyzed. In the present study, intracellular self-assembly of key components of cellulosomes independent of Ca2+ was achieved, and DocA-S3 was applied to the assembly of key enzymes of L-lysine biosynthesis, in which DapA and DapB intracellular assembly increased L-lysine accumulation by 29.8% when compared with the control strains, providing a new strategy for improving the intracellular self-assembly of cellulosomes and amino acid fermentation efficiency.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"82 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140655170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-04-25DOI: 10.3390/fermentation10050226
Suzane Meriely da Silva Duarte, Allysson Kayron de Carvalho Silva, Katia Regina Assunção Borges, Carolina Borges Cordeiro, Fernanda Jeniffer Lindoso Lima, Marcos Antônio Custódio Neto da Silva, Marcelo de Souza Andrade, Maria do Desterro Soares Brandão Nascimento
Cervical cancer is caused by a persistent and high-grade infection. It is caused by the Human Papillomavirus (HPV), which, when entering cervical cells, alters their physiology and generates serious lesions. HPV 18 is among those most involved in carcinogenesis in this region, but there are still no drug treatments that cause cure or total remission of lesions caused by HPV. It is known that L-asparaginase is an amidohydrolase, which plays a significant role in the pharmaceutical industry, particularly in the treatment of specific cancers. Due to its antitumor properties, some studies have demonstrated its cytotoxic effect against cervical cancer cells. However, the commercial version of this enzyme has side effects, such as hypersensitivity, allergic reactions, and silent inactivation due to the formation of antibodies. To mitigate these adverse effects, several alternatives have been explored, including the use of L-asparaginase from other microbiological sources, which is the case with the use of the fungus Aspergillus niger, a high producer of L-asparaginase. The study investigated the influence of the type of fermentation, precipitant, purification, characterization, and in vitro cytotoxicity of L-asparaginase. The results revealed that semisolid fermentation produced higher enzymatic activity and protein concentration of A. niger. The characterized enzyme showed excellent stability at pH 9.0, temperature of 50 °C, resistance to surfactants and metallic ions, and an increase in enzymatic activity with the organic solvent ethanol. Furthermore, it exhibited low cytotoxicity in GM and RAW cells and significant cytotoxicity in HeLa cells. These findings indicate that L-asparaginase derived from A. niger may be a promising alternative for pharmaceutical production. Its attributes, including stability, activity, and low toxicity in healthy cells, suggest that this modified enzyme could overcome challenges associated with antitumor therapy.
宫颈癌是由持续的高级别感染引起的。它是由人类乳头状瘤病毒（HPV）引起的，HPV 进入宫颈细胞后会改变其生理结构并产生严重病变。人乳头瘤病毒 18 是该地区致癌作用最强的病毒之一，但目前还没有药物能治愈或完全缓解由人乳头瘤病毒引起的病变。众所周知，L-天冬酰胺酶是一种酰胺水解酶，在制药业，特别是在治疗特定癌症方面发挥着重要作用。由于其抗肿瘤特性，一些研究已经证明了它对宫颈癌细胞的细胞毒性作用。然而，这种酶的商业版本有副作用，如过敏、过敏反应，以及因形成抗体而导致的无声失活。为了减轻这些副作用，人们探索了几种替代方法，包括使用其他微生物来源的 L-天冬酰胺酶，例如使用黑曲霉这种 L-天冬酰胺酶产量很高的真菌。该研究调查了 L-天冬酰胺酶的发酵类型、沉淀剂、纯化、表征和体外细胞毒性的影响。结果表明，半固态发酵能产生更高的黑木耳酶活性和蛋白质浓度。所表征的酶在 pH 值为 9.0、温度为 50 ℃ 的条件下表现出极佳的稳定性，对表面活性剂和金属离子具有抗性，在有机溶剂乙醇的作用下酶活性也有所提高。此外，它在 GM 和 RAW 细胞中表现出较低的细胞毒性，而在 HeLa 细胞中则表现出显著的细胞毒性。这些研究结果表明，从黑木耳中提取的 L-天冬酰胺酶可能是一种很有前景的制药替代品。它的特性，包括稳定性、活性和对健康细胞的低毒性，表明这种改良酶可以克服与抗肿瘤治疗相关的挑战。
{"title":"Production, Characterization Purification, and Antitumor Activity of L-Asparaginase from Aspergillus niger","authors":"Suzane Meriely da Silva Duarte, Allysson Kayron de Carvalho Silva, Katia Regina Assunção Borges, Carolina Borges Cordeiro, Fernanda Jeniffer Lindoso Lima, Marcos Antônio Custódio Neto da Silva, Marcelo de Souza Andrade, Maria do Desterro Soares Brandão Nascimento","doi":"10.3390/fermentation10050226","DOIUrl":"https://doi.org/10.3390/fermentation10050226","url":null,"abstract":"Cervical cancer is caused by a persistent and high-grade infection. It is caused by the Human Papillomavirus (HPV), which, when entering cervical cells, alters their physiology and generates serious lesions. HPV 18 is among those most involved in carcinogenesis in this region, but there are still no drug treatments that cause cure or total remission of lesions caused by HPV. It is known that L-asparaginase is an amidohydrolase, which plays a significant role in the pharmaceutical industry, particularly in the treatment of specific cancers. Due to its antitumor properties, some studies have demonstrated its cytotoxic effect against cervical cancer cells. However, the commercial version of this enzyme has side effects, such as hypersensitivity, allergic reactions, and silent inactivation due to the formation of antibodies. To mitigate these adverse effects, several alternatives have been explored, including the use of L-asparaginase from other microbiological sources, which is the case with the use of the fungus Aspergillus niger, a high producer of L-asparaginase. The study investigated the influence of the type of fermentation, precipitant, purification, characterization, and in vitro cytotoxicity of L-asparaginase. The results revealed that semisolid fermentation produced higher enzymatic activity and protein concentration of A. niger. The characterized enzyme showed excellent stability at pH 9.0, temperature of 50 °C, resistance to surfactants and metallic ions, and an increase in enzymatic activity with the organic solvent ethanol. Furthermore, it exhibited low cytotoxicity in GM and RAW cells and significant cytotoxicity in HeLa cells. These findings indicate that L-asparaginase derived from A. niger may be a promising alternative for pharmaceutical production. Its attributes, including stability, activity, and low toxicity in healthy cells, suggest that this modified enzyme could overcome challenges associated with antitumor therapy.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"30 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140658408","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-04-25DOI: 10.3390/fermentation10050228
F. P. Spada, S. D. de Alencar, Eduardo Purgatto, Stanislau Bogusz Junior
Cocoa is the third most important global agricultural export commodity. However, because it is a crop sensitive to climatic change, there has been an active search for cocoa substitutes worldwide. Roasted jackfruit seeds were previously described as having a chocolate aroma and are affordable and accessible. In this study, we characterized and identified by SPME-GC-O and SPME-GC-MS the aroma profile of cocoa-based beverages formulated with jackfruit seed flour as a natural cocoa substitute. Our analysis tentatively identified 71 odor-active aroma descriptors with some similarities between formulations. Overall, 15 odor-active aromas were present in all beverages. The formulation containing only cocoa/chocolate showed the following aroma descriptors: cocoa, hazelnut, peanut butter, earthy, and roast, which are mostly related to the presence of 2,3-dimethylpyrazine and 2,3-diethyl-5-methylpyrazine. The fermented beverage had a content of complex pyrazines such as 2,3,5-trimethyl-6-isopentylpyrazine and methylpropylpyrazine. Our data indicated that both the control and fermented beverages showed a similar aromatic profile, mainly earthy, pyrazine, and chocolate. Qualitative similarities in the pyrazine content were observed between the fermented jackfruit seed flour and cocoa beverages. In conclusion, fermented jackfruit seed flour can be incorporated into cocoa-based beverages as a natural chocolate substitute, offering the potential to elevate the chocolate aroma.
{"title":"GC-Olfactometric Analysis as a Tool for Comprehensive Characterization of Aromatic Profiles in Cocoa-Based Beverages with a Natural Chocolate Substitute","authors":"F. P. Spada, S. D. de Alencar, Eduardo Purgatto, Stanislau Bogusz Junior","doi":"10.3390/fermentation10050228","DOIUrl":"https://doi.org/10.3390/fermentation10050228","url":null,"abstract":"Cocoa is the third most important global agricultural export commodity. However, because it is a crop sensitive to climatic change, there has been an active search for cocoa substitutes worldwide. Roasted jackfruit seeds were previously described as having a chocolate aroma and are affordable and accessible. In this study, we characterized and identified by SPME-GC-O and SPME-GC-MS the aroma profile of cocoa-based beverages formulated with jackfruit seed flour as a natural cocoa substitute. Our analysis tentatively identified 71 odor-active aroma descriptors with some similarities between formulations. Overall, 15 odor-active aromas were present in all beverages. The formulation containing only cocoa/chocolate showed the following aroma descriptors: cocoa, hazelnut, peanut butter, earthy, and roast, which are mostly related to the presence of 2,3-dimethylpyrazine and 2,3-diethyl-5-methylpyrazine. The fermented beverage had a content of complex pyrazines such as 2,3,5-trimethyl-6-isopentylpyrazine and methylpropylpyrazine. Our data indicated that both the control and fermented beverages showed a similar aromatic profile, mainly earthy, pyrazine, and chocolate. Qualitative similarities in the pyrazine content were observed between the fermented jackfruit seed flour and cocoa beverages. In conclusion, fermented jackfruit seed flour can be incorporated into cocoa-based beverages as a natural chocolate substitute, offering the potential to elevate the chocolate aroma.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"53 48","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140656477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-04-25DOI: 10.3390/fermentation10050227
O. Ikusika, O. Akinmoladun, C. T. Mpendulo
The abundance of fruit waste from the food industry and wineries, particularly peels, seeds, and other fruit pomace throughout the year, could lead to health and environmental hazards if not channelled into productive areas. Improving or transforming these waste products for better use in other vital sectors could be achieved via solid-state fermentation (SSF) since most waste products are solid. One such productive and important area is the feeding of livestock, which will guarantee millennium food security goals for many nations of the world. The nutritional and antioxidant composition of abundantly available fruit pomace and agro-industrial byproducts could be improved via solid-state fermentation for overall livestock productivity. They contain substantial dietary fibre, protein, and phenolic compounds; hence, improving them via fermentation could serve the livestock industry in dual capacities, including nutraceutical and conventional feedstuff. This review seeks to provide reinforcing evidence on the applicability and impact of fruit pomaces on livestock nutrition. The significant nutrient improvements, beneficial outcomes in feeding trials, and inconsistencies or areas of research gap were also explored.
{"title":"Enhancement of the Nutritional Composition and Antioxidant Activities of Fruit Pomaces and Agro-Industrial Byproducts through Solid-State Fermentation for Livestock Nutrition: A Review","authors":"O. Ikusika, O. Akinmoladun, C. T. Mpendulo","doi":"10.3390/fermentation10050227","DOIUrl":"https://doi.org/10.3390/fermentation10050227","url":null,"abstract":"The abundance of fruit waste from the food industry and wineries, particularly peels, seeds, and other fruit pomace throughout the year, could lead to health and environmental hazards if not channelled into productive areas. Improving or transforming these waste products for better use in other vital sectors could be achieved via solid-state fermentation (SSF) since most waste products are solid. One such productive and important area is the feeding of livestock, which will guarantee millennium food security goals for many nations of the world. The nutritional and antioxidant composition of abundantly available fruit pomace and agro-industrial byproducts could be improved via solid-state fermentation for overall livestock productivity. They contain substantial dietary fibre, protein, and phenolic compounds; hence, improving them via fermentation could serve the livestock industry in dual capacities, including nutraceutical and conventional feedstuff. This review seeks to provide reinforcing evidence on the applicability and impact of fruit pomaces on livestock nutrition. The significant nutrient improvements, beneficial outcomes in feeding trials, and inconsistencies or areas of research gap were also explored.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"48 21","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140657008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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