文献类型：期刊文献

英文题名：Production of L-phenylalanine from trans-cinnamic acids by high-level expression of phenylalanine ammonia lyase gene from Rhodosporidium toruloides in Escherichia coli

作者：Jia, Shi Ru[1];Cui, Jian Dong[2];Li, Yan[2];Sun, Ai You[3]

机构：[1]Tianjin Univ Sci & Technol, Tianjin Key Lab Ind Microbiol, Tai Dev Area, Tianjin 300457, Peoples R China;[2]Hebei Univ Sci & Technol, Coll Bioengn, Shijiazhuang 050018, Peoples R China;[3]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2008

卷号：42

期号：3

起止页码：193

外文期刊名：BIOCHEMICAL ENGINEERING JOURNAL

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000260283800001)】；

语种：英文

外文关键词：Phenylalanine ammonia-lyase; Recombinant DNA; Combine promoter; Enzyme activity; Fermentation; Bioconversion

摘要：A combined promoter expression vector pBV-PAL for high-level expression of phenylalanine ammonia lyase gene of Rhodosporidium toruloides was constructed. Pal gene was cloned and inserted into the region between Sall and PstI restriction sites of expression vector pBV220 (containing P(L)P(R) promoter) to obtain recombinant expression vector pBV220-PAL. The tac promoter obtained from the plasmid pKtac was inserted into the expression vector pBV220-PAL to construct expression vector pBV-PAL. The recombinant plasmid pBV220-PAL and pBV-PAL were introduced into Escherichia coli JM109 by transformation. The result showed that the transformant E. coli JM109 (pBV-PAL) gave a much higher PAL activity than that transformant E coli JM109 (pBV220-PAL). Recombinant PAL expression level of the transformant JM109 (pBV-PAL) was about 9.6% of total cellular protein, specific enzyme activity was 2.3-fold higher than that of the transformant JM109 (pBV220-PAL), reached 35 U/g (dry cells weight, DCW). PAL specific activity of 123 U/g (DCW) could be achieved in a 5-l fermentor. 80.5% conversion rate of trans-cinnamic acid to L-phenylalanine and 5.12 g/l L-phenylalanine were obtained after 3 h bioconversion using the transformant JM109 (pBV-PAL). The recombinant strain JM109 containing the combined promoter expression vector pBV-PAL was shown to be effective and practical to product L-phenylalanine. (C) 2008 Elsevier B.V. All rights reserved.