Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxii
4-Hydroxy-2,5-dimethyl-3 (2H)-furanone (HDMF) is widely used in the food industry as a spice and flavoring agent with high market demand. In this study, fructose-1,6-bisphosphate aldolase (FBA) and triose phosphate isomerase (TPI) were overexpressed in Zygosaccharomyces rouxii in the form of single...
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Frontiers Media S.A.
2024-03-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1366021/full |
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| author | Yanhong Wang Wei Liu Jingyao Chen Zhijiang Li Yijia Hu Zixiang Fan Liangyuan Yan Jiahui Liu Yuao Zhou Wei Jiang Haiying Rui Lingyan Dai |
| author_facet | Yanhong Wang Wei Liu Jingyao Chen Zhijiang Li Yijia Hu Zixiang Fan Liangyuan Yan Jiahui Liu Yuao Zhou Wei Jiang Haiying Rui Lingyan Dai |
| author_sort | Yanhong Wang |
| collection | DOAJ |
| description | 4-Hydroxy-2,5-dimethyl-3 (2H)-furanone (HDMF) is widely used in the food industry as a spice and flavoring agent with high market demand. In this study, fructose-1,6-bisphosphate aldolase (FBA) and triose phosphate isomerase (TPI) were overexpressed in Zygosaccharomyces rouxii in the form of single and double genes, respectively, via electroporation. High-yield HDMF-engineered yeast strains were constructed by combining the analysis of gene expression levels obtained by real-time fluorescence quantitative PCR technology and HDMF production measured by HPLC. The results showed that there was a significant positive correlation between the production of HDMF and the expression levels of the FBA and TPI genes in yeast; the expression levels of the FBA and TPI genes were also positively correlated (p < 0.05). Compared with the wild type (WT), the engineered strains F10-D, T17-D, and TF15-A showed marked increases in HDMF production and FBA and TPI gene expression (p < 0.05) and exhibited great genetic stability with no obvious differences in biomass or colony morphology. In addition, the exogenous addition of d-fructose promoted the growth of Z. rouxii. Among the engineered strains, when fermented in YPD media supplemented with d-fructose for 5 days, TF15-A (overexpressing the FBA and TPI genes) generated the highest HDMF production of 13.39 mg/L, which is 1.91 times greater than that of the wild-type strain. The results above indicated that FBA and TPI, which are key enzymes involved in the process of HDMF biosynthesis by Z. rouxii, positively regulate the synthesis of HDMF at the transcriptional level. d-fructose can be used as a precursor for the biosynthesis of HDMF by engineered yeast in industrial production. |
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| format | Article |
| id | doaj.art-cbd60ce90c2e48f19a01000811152053 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-24T22:24:04Z |
| publishDate | 2024-03-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-cbd60ce90c2e48f19a010008111520532024-03-20T04:59:12ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-03-011510.3389/fmicb.2024.13660211366021Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxiiYanhong Wang0Wei Liu1Jingyao Chen2Zhijiang Li3Yijia Hu4Zixiang Fan5Liangyuan Yan6Jiahui Liu7Yuao Zhou8Wei Jiang9Haiying Rui10Lingyan Dai11Heilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, ChinaHeilongjiang Agricultural Economy Vocational College, Mudanjiang, ChinaHeilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, ChinaCollege of Food Science, Heilongjiang Bayi Agricultural University, Daqing, ChinaCollege of Food Science, Heilongjiang Bayi Agricultural University, Daqing, ChinaHeilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, ChinaHeilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, ChinaHeilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, ChinaHeilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, ChinaHeilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, ChinaDaqing Branch of Heilongjiang Academy of Agricultural Sciences, Daqing, ChinaHeilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, China4-Hydroxy-2,5-dimethyl-3 (2H)-furanone (HDMF) is widely used in the food industry as a spice and flavoring agent with high market demand. In this study, fructose-1,6-bisphosphate aldolase (FBA) and triose phosphate isomerase (TPI) were overexpressed in Zygosaccharomyces rouxii in the form of single and double genes, respectively, via electroporation. High-yield HDMF-engineered yeast strains were constructed by combining the analysis of gene expression levels obtained by real-time fluorescence quantitative PCR technology and HDMF production measured by HPLC. The results showed that there was a significant positive correlation between the production of HDMF and the expression levels of the FBA and TPI genes in yeast; the expression levels of the FBA and TPI genes were also positively correlated (p < 0.05). Compared with the wild type (WT), the engineered strains F10-D, T17-D, and TF15-A showed marked increases in HDMF production and FBA and TPI gene expression (p < 0.05) and exhibited great genetic stability with no obvious differences in biomass or colony morphology. In addition, the exogenous addition of d-fructose promoted the growth of Z. rouxii. Among the engineered strains, when fermented in YPD media supplemented with d-fructose for 5 days, TF15-A (overexpressing the FBA and TPI genes) generated the highest HDMF production of 13.39 mg/L, which is 1.91 times greater than that of the wild-type strain. The results above indicated that FBA and TPI, which are key enzymes involved in the process of HDMF biosynthesis by Z. rouxii, positively regulate the synthesis of HDMF at the transcriptional level. d-fructose can be used as a precursor for the biosynthesis of HDMF by engineered yeast in industrial production.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1366021/full4-Hydroxy-2,5-dimethyl-3 (2H)-furanone (HDMF)Zygosaccharomyces rouxiifructose-1,6-bisphosphate aldolase (FBA)triose phosphate isomerase (TPI)engineered yeast |
| spellingShingle | Yanhong Wang Wei Liu Jingyao Chen Zhijiang Li Yijia Hu Zixiang Fan Liangyuan Yan Jiahui Liu Yuao Zhou Wei Jiang Haiying Rui Lingyan Dai Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxii Frontiers in Microbiology 4-Hydroxy-2,5-dimethyl-3 (2H)-furanone (HDMF) Zygosaccharomyces rouxii fructose-1,6-bisphosphate aldolase (FBA) triose phosphate isomerase (TPI) engineered yeast |
| title | Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxii |
| title_full | Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxii |
| title_fullStr | Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxii |
| title_full_unstemmed | Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxii |
| title_short | Overexpression of the FBA and TPI genes promotes high production of HDMF in Zygosaccharomyces rouxii |
| title_sort | overexpression of the fba and tpi genes promotes high production of hdmf in zygosaccharomyces rouxii |
| topic | 4-Hydroxy-2,5-dimethyl-3 (2H)-furanone (HDMF) Zygosaccharomyces rouxii fructose-1,6-bisphosphate aldolase (FBA) triose phosphate isomerase (TPI) engineered yeast |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1366021/full |
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