{"title":"代谢工程大肠杆菌从营养性二糖蔗糖生物合成非营养性单糖 d-阿洛糖","authors":"Ling-Jie Zheng, Wei-Xiang Chen, Shang-He Zheng, Irfan Ullah, Hui-Dong Zheng, Li-Hai Fan, Qiang Guo","doi":"10.1002/bit.28842","DOIUrl":null,"url":null,"abstract":"<p>Sucrose is a commonly utilized nutritive sweetener in food and beverages due to its abundance in nature and low production costs. However, excessive intake of sucrose increases the risk of metabolic disorders, including diabetes and obesity. Therefore, there is a growing demand for the development of nonnutritive sweeteners with almost no calories. <span>d</span>-Allulose is an ultra-low-calorie, rare six-carbon monosaccharide with high sweetness, making it an ideal alternative to sucrose. In this study, we developed a cell factory for <span>d</span>-allulose production from sucrose using <i>Escherichia coli</i> JM109 (DE3) as a chassis host. The genes <i>cscA</i>, <i>cscB</i>, <i>cscK</i>, <i>alsE</i>, and <i>a6PP</i> were co-expressed for the construction of the synthesis pathway. Then, the introduction of <i>ptsG-F</i> and knockout of <i>ptsG</i>, <i>fruA</i>, <i>ptsI</i>, and <i>ptsH</i> to reprogram sugar transport pathways resulted in an improvement in substrate utilization. Next, the carbon fluxes of the Embden-Meyerhof-Parnas and the pentose phosphate pathways were regulated by the inactivation of <i>pfkA</i> and <i>zwf</i>, achieving an increase in <span>d</span>-allulose titer and yield of 154.2% and 161.1%, respectively. Finally, scaled-up fermentation was performed in a 5 L fermenter. The titer of <span>d</span>-allulose reached 11.15 g/L, with a yield of 0.208 g/g on sucrose.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 12","pages":"3684-3693"},"PeriodicalIF":3.5000,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Biosynthesis of nonnutritive monosaccharide d-allulose by metabolically engineered Escherichia coli from nutritive disaccharide sucrose\",\"authors\":\"Ling-Jie Zheng, Wei-Xiang Chen, Shang-He Zheng, Irfan Ullah, Hui-Dong Zheng, Li-Hai Fan, Qiang Guo\",\"doi\":\"10.1002/bit.28842\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Sucrose is a commonly utilized nutritive sweetener in food and beverages due to its abundance in nature and low production costs. However, excessive intake of sucrose increases the risk of metabolic disorders, including diabetes and obesity. Therefore, there is a growing demand for the development of nonnutritive sweeteners with almost no calories. <span>d</span>-Allulose is an ultra-low-calorie, rare six-carbon monosaccharide with high sweetness, making it an ideal alternative to sucrose. In this study, we developed a cell factory for <span>d</span>-allulose production from sucrose using <i>Escherichia coli</i> JM109 (DE3) as a chassis host. The genes <i>cscA</i>, <i>cscB</i>, <i>cscK</i>, <i>alsE</i>, and <i>a6PP</i> were co-expressed for the construction of the synthesis pathway. Then, the introduction of <i>ptsG-F</i> and knockout of <i>ptsG</i>, <i>fruA</i>, <i>ptsI</i>, and <i>ptsH</i> to reprogram sugar transport pathways resulted in an improvement in substrate utilization. Next, the carbon fluxes of the Embden-Meyerhof-Parnas and the pentose phosphate pathways were regulated by the inactivation of <i>pfkA</i> and <i>zwf</i>, achieving an increase in <span>d</span>-allulose titer and yield of 154.2% and 161.1%, respectively. Finally, scaled-up fermentation was performed in a 5 L fermenter. The titer of <span>d</span>-allulose reached 11.15 g/L, with a yield of 0.208 g/g on sucrose.</p>\",\"PeriodicalId\":9168,\"journal\":{\"name\":\"Biotechnology and Bioengineering\",\"volume\":\"121 12\",\"pages\":\"3684-3693\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2024-09-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biotechnology and Bioengineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/bit.28842\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology and Bioengineering","FirstCategoryId":"5","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/bit.28842","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Biosynthesis of nonnutritive monosaccharide d-allulose by metabolically engineered Escherichia coli from nutritive disaccharide sucrose
Sucrose is a commonly utilized nutritive sweetener in food and beverages due to its abundance in nature and low production costs. However, excessive intake of sucrose increases the risk of metabolic disorders, including diabetes and obesity. Therefore, there is a growing demand for the development of nonnutritive sweeteners with almost no calories. d-Allulose is an ultra-low-calorie, rare six-carbon monosaccharide with high sweetness, making it an ideal alternative to sucrose. In this study, we developed a cell factory for d-allulose production from sucrose using Escherichia coli JM109 (DE3) as a chassis host. The genes cscA, cscB, cscK, alsE, and a6PP were co-expressed for the construction of the synthesis pathway. Then, the introduction of ptsG-F and knockout of ptsG, fruA, ptsI, and ptsH to reprogram sugar transport pathways resulted in an improvement in substrate utilization. Next, the carbon fluxes of the Embden-Meyerhof-Parnas and the pentose phosphate pathways were regulated by the inactivation of pfkA and zwf, achieving an increase in d-allulose titer and yield of 154.2% and 161.1%, respectively. Finally, scaled-up fermentation was performed in a 5 L fermenter. The titer of d-allulose reached 11.15 g/L, with a yield of 0.208 g/g on sucrose.
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