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| Research on the high-level expression of proline hydroxylase induced by lactose |
| Ma Lehui， Li Cunhui |
| Hebei Bolunte Pharmaceutical Corporation Ltd.， Shijiazhuang 050000, China |
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Abstract The expression of proline hydroxylase in escherichia coli induced by lactose and the biotransformation of L-proline were studied. The results show that first the seed liquid was inoculated in the 0.8% lactose fermentation medium according to the proportion of 1%, and at 28 ℃ and 140 r/m， the bacteria was cultured for about 2 hours. Then under the condition that the wavelength of the spectrophotometer is 600 nm and the OD value is 0.6 ~ 0.8, the culture liquid was induced and cultured for 13 hours, and the bacteria was collected by centrifugation. Finally the conversion solution was added to the bacteria according to the ratio of 1 g∶100 mL ( biomass volume∶conversion solution ), and L-proline was efficiently transformed into L-hydroxyproline at 28 ℃, which conversion rate reached 98.6%. This study lays a foundation for the industrialization of L-proline biotransformation.
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| [ 1 ] 刘合栋. 高产反式-4-羟脯氨酸重组大肠杆菌的构建和发酵优化[ D ]. 无锡：江南大学，2013：1 - 57．
[ 2 ] Hara R, Kino K. Characterization of novel 2-oxoglutarate dependent dioxygenases converting L- proline to cis -4- hydroxy -L- proline[ J ]. Biochemical and Biophysical Reserch Communications，2009，379: 882 - 886.
[3 ] 高焕春，李文英. 从骨胶水解液中分离L-羟脯氨酸和L-脯氨酸[ J ]. 天津轻工业学院学报，1997（ 2 ）：33 - 37.
[ 4 ] 吕红线. 从骨胶中制备L-羟脯氨酸的研究[ J ]. 山东轻工业学院学报：自然科学版，1997，11（ 4 ）：53 - 56.
[ 5 ] 李 娟，陈舜胜. 从明胶水解液中分离制备L-羟脯氨酸和L-脯氨酸[ J ]. 现代食品科技，2006（ 4 ）：151 - 152．
[ 6 ] 袁春伟，何艳春，张胜利，等. 重组大肠杆菌 BL21（pUC19-Hyp）产羟脯氨酸的补料分批培养[ J ]. 生物加工过程，2014，12（ 4 ）：43 - 48．
[ 7 ] Shibasaki T, Mori H, Ozaki A. Enzymatic production of trans - 4 - Hydroxy-L-proline by regio-and stereospecific hydroxylation of L -Proline[ J ]. Bioscience, Biotechnology and Biochemistry, 2000, 64（ 4 ）：746 - 750．
[ 8 ] 吴一凡，张双全，高秀玉，等. 乳糖诱导pET载体表达重组蛋白的研究[ J ]. 南京师大学报，2002，25（ 1 ）：89 - 93．
[ 9 ] Jobe A, Bourgeois S. lacrepressor-operator interaction: VI. The natural inducer of the lacoperon[ J ]. J Mol Biol，1972，69（ 3 ）: 397 - 404．
[ 10 ] Muller-Hill B, Rickenberg HV, Wallenfels K. Specificity of the induction of the enzymes of the lac operon in Es-cherichia coli[ J ]. J Mol Biol, 1964, 10（ 2 ）：303 - 318．
[ 11 ] 杨海麟，王长城，张 玲，等. 高密度培养重组E-coli产胆固醇氧化酶的乳糖诱导策略[ J ]. 食品工业科技，2011，2：162 - 165．
[ 12 ] 田文静，罗学刚，吕丽慧，等. 乳糖诱导瑞替普酶在大肠杆菌中的可溶性表达[ J ]. 中国生化药物杂志，2011，32（ 5 ）：341 - 348．
[ 13 ] 李兆鹏，张 栩，徐 斌，等. 重组大肠杆菌高密度发酵中乳糖诱导表达hBLyS[ J ]. 过程工程学报，2005，5（ 4 ）：446 - 449. |
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