{"title":"培养重组大肠杆菌,获得高细胞密度和高水平的青霉素G酰化酶活性。","authors":"Y C Liu, L C Liao, W T Wu","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>A mutant strain of E. coli EP1 harbouring pGL-5 was employed to develop a process for producing penicillin G acylase (PGA). In comparison with different carbon sources in the medium, it was found that the specific levels of PGA activity obtained in the glucose medium were the lowest. which was likely due to catabolic repression. Phenylacetic acid (PAA) was previously reported to be an regulatory inducer for PGA production, whereas in this study, the addition of PAA repressed both cell growth and enzyme expression. In a fed-batch culture, the increase of specific PGA activity followed the pattern of the cell concentration during the early to middle cell growth phase. With application of pure oxygen aeration and an appropriate medium design, the cell concentration reached 162 (g wet weight/l), which was 2.4 times higher compared to that of the original operation, and a specific PGA activity of 37 (IU/g wet weight) was achieved after 12 h of cultivation.</p>","PeriodicalId":20569,"journal":{"name":"Proceedings of the National Science Council, Republic of China. Part B, Life sciences","volume":"24 4","pages":"156-60"},"PeriodicalIF":0.0000,"publicationDate":"2000-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Cultivation of recombinant Escherichia coli to achieve high cell density with a high level of penicillin G acylase activity.\",\"authors\":\"Y C Liu, L C Liao, W T Wu\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>A mutant strain of E. coli EP1 harbouring pGL-5 was employed to develop a process for producing penicillin G acylase (PGA). In comparison with different carbon sources in the medium, it was found that the specific levels of PGA activity obtained in the glucose medium were the lowest. which was likely due to catabolic repression. Phenylacetic acid (PAA) was previously reported to be an regulatory inducer for PGA production, whereas in this study, the addition of PAA repressed both cell growth and enzyme expression. In a fed-batch culture, the increase of specific PGA activity followed the pattern of the cell concentration during the early to middle cell growth phase. With application of pure oxygen aeration and an appropriate medium design, the cell concentration reached 162 (g wet weight/l), which was 2.4 times higher compared to that of the original operation, and a specific PGA activity of 37 (IU/g wet weight) was achieved after 12 h of cultivation.</p>\",\"PeriodicalId\":20569,\"journal\":{\"name\":\"Proceedings of the National Science Council, Republic of China. Part B, Life sciences\",\"volume\":\"24 4\",\"pages\":\"156-60\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2000-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Proceedings of the National Science Council, Republic of China. Part B, Life sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Proceedings of the National Science Council, Republic of China. Part B, Life sciences","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Cultivation of recombinant Escherichia coli to achieve high cell density with a high level of penicillin G acylase activity.
A mutant strain of E. coli EP1 harbouring pGL-5 was employed to develop a process for producing penicillin G acylase (PGA). In comparison with different carbon sources in the medium, it was found that the specific levels of PGA activity obtained in the glucose medium were the lowest. which was likely due to catabolic repression. Phenylacetic acid (PAA) was previously reported to be an regulatory inducer for PGA production, whereas in this study, the addition of PAA repressed both cell growth and enzyme expression. In a fed-batch culture, the increase of specific PGA activity followed the pattern of the cell concentration during the early to middle cell growth phase. With application of pure oxygen aeration and an appropriate medium design, the cell concentration reached 162 (g wet weight/l), which was 2.4 times higher compared to that of the original operation, and a specific PGA activity of 37 (IU/g wet weight) was achieved after 12 h of cultivation.