Abstract
3-Hydroxypropionic acid (3-HP) is an important platform chemical in organic synthesis. Traditionally, 3-HP was produced by chemical methods and fermentation process. In this work, a novel enzymatic method was developed for green synthesis of 3-HP. A yeast strain harboring nitrile-hydrolyzing enzyme was newly isolated from environmental samples using 3-hydroxypropionitrile (3-HPN) as the sole nitrogen source. It was identified to be Meyerozyma guilliermondii CGMCC12935 by sequencing of the 18S ribosomal DNA and internal transcribed spacer, together with analysis of the morphology characteristics. The catalytic properties of M. guilliermondii CGMCC12935 resting cells were determined, and the optimum activity was achieved at 55 °C and pH 7.5. The enzyme showed broad substrate specificity towards nitriles, especially 3-HPN, aminoacetonitrile and 3-cyanopyridine. The presence of Ag+, Pb2+ and excess substrate inhibited the enzyme activity, whereas 5% (v/v) ethyl acetate had a positive effect on the enzyme activity. M. guilliermondii CGMCC12935 resting cells by addition of 3% glucose could thoroughly hydrolyze 500 mM 3-HPN into 3-HP within 100 h and the maximal accumulative production of 3-HP reached 216.33 mM, which was over twofolds than the control group with no additional glucose. And this work would lay the foundation for biological production of 3-HP in industry.
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Acknowledgements
The work was supported by the National Natural Science Foundation of China (No. 21406088), the Natural Science Foundation of Jiangsu Province (No. BK20140133), and the Key Project of Science and Technology Development Program of Shandong Province (No. 2016ZDJS07B08).
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449_2017_1754_MOESM1_ESM.tif
Phylogenetic tree of M. guilliermondii and related strains based on ITS sequences, constructed by the neighbor-joining method. The accession numbers of these strains were given in brackets. Bootstrap values were based on 1,000 replicates (TIF 59 KB)
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Zhang, Q., Gong, JS., Dong, TT. et al. Nitrile-hydrolyzing enzyme from Meyerozyma guilliermondii and its potential in biosynthesis of 3-hydroxypropionic acid. Bioprocess Biosyst Eng 40, 901–910 (2017). https://doi.org/10.1007/s00449-017-1754-6
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DOI: https://doi.org/10.1007/s00449-017-1754-6