期刊
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
卷 129, 期 -, 页码 37-42出版社
ELSEVIER
DOI: 10.1016/j.molcatb.2016.03.011
关键词
Nitrilase; Immobilization; 3-Hydroxypropionic acid; Hydrolysis; Enzyme catalysis
资金
- National Natural Science Foundation of China [21572261]
- Key Project of Tianjin Municipal Science and Technology Support Program [15ZCZDSY00490]
- Youth Innovation Promotion Association, CAS
3-Hydroxypropionic acid (3-HP) is an important platform chemical for organic synthesis and high performance polymers. Despite a wealth of reports related to 3-HP biosynthesis in microorganisms, its industrial application still requires further research because of low titer and productivity. Herein an effective enzymatic method for the synthesis of 3-HP was achieved by using free or immobilized recombinant Escherichia coli BL21(DE3) cells harboring a nitrilase gene from environmental sample (NIT190). Under the optimal conditions (100 mmol/LTris-HCl buffer, pH 8.0, 30 degrees C), the maximum substrate concentration which could be completely hydrolyzed by using free cells within 24h was 4.5 mol/L (319.5 g/L). Furthermore, immobilization of the whole cells enhanced their substrate tolerance (up to 7.0 mol/L), stability, and reusability. The immobilized cells could be reused for up to 30 batches, and 70% of enzyme activity was retained after 74 batches in distilled water. The titer (184.7 g/L) and productivity (36.9 g/(Lh)) were obtained by isolation and purification of 3-HP from the first 30 batches. These results demonstrate that the immobilized cells have potential industrial application for the synthesis of 3-HP. (C) 2016 Elsevier B.V. All rights reserved.
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