Identification and Mutagenesis of a New Isolated Strain Bacillus sp. B26 for Producing(R)-α-Hydroxyphenylacetic Acid

›› 2011, Vol. 19 ›› Issue (4): 636-643.

• BIOTECHNOLOGY AND BIOENGINEERING • Previous Articles Next Articles

Identification and Mutagenesis of a New Isolated Strain Bacillus sp. B26 for Producing(R)-α-Hydroxyphenylacetic Acid

CHEN Bingmei¹, XU Xiaoping¹, HOU Zhiguo¹, LI Zhongqin², RUAN Wenbing¹

1. College of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350108, China;
2. College of Fisheries, Jimei University, Xiamen 361021, China

Received:2011-01-06 Revised:2011-04-18 Online:2011-08-28 Published:2011-08-28
Supported by:
Supported by the Specialized Research Fund for the Doctoral Program of Higher Education of China (20103514110002);the Science and Technology Key Project of Fujian Province (2009N0046)

Identification and Mutagenesis of a New Isolated Strain Bacillus sp. B26 for Producing(R)-α-Hydroxyphenylacetic Acid

陈秉梅¹, 许小平¹, 侯志国¹, 李忠琴², 阮文兵¹

1. College of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350108, China;
2. College of Fisheries, Jimei University, Xiamen 361021, China

通讯作者: XU Xiaoping,E-mail:xu@fzu.edu.cn
基金资助:
Supported by the Specialized Research Fund for the Doctoral Program of Higher Education of China (20103514110002);the Science and Technology Key Project of Fujian Province (2009N0046)

Abstract

Abstract: A bacterium strain B26 capable of producing(R)-α-hydroxyphenylacetic acid [(R)-HPA](yield, 47.5%;enantiomeric excess, 99.1%) from phenylglyoxylic acid(PGA) with high optical purity was isolated and identified as Bacillus sp.B26 by 16S rDNA(ribosomal DNA) sequencing.Phylogenic analysis showed that the strain was most similar to Bacillus sp.enrichment culture clone SYW5(FJ601635.1) and Bacillus cereus strain FM-4(EU794727.1).Efforts were made to further improve HPA-production and PGA-tolerance by UV irradiation and UV-LiCl cooperative mutagenesis.Among viable mutants, B.sp.UV-38 and B.sp.ULi-11 exhibited better productivities than the wild type.Comparisons of HPA production and time course among wild strain and two mutants showed that B.sp.ULi-11 was more competent than B.sp.UV-38.HPA production was increased by 39.1% with B.sp.ULi-11(yield, 65.4%) compared to that with B.sp.B26(yield, 47.0%) when cultured in fermentation broth(pH 7.2) at 32℃ with an agitation speed of 180 r·min^-1 and PGA final concentration of 15 mmol·L^-1 for 25 h.

Key words: (R)-α-hydroxyphenylacetic acid, biotransformation, phylogenetic analysis, Bacillus, phenylglyoxylic acid, UV mutagenesis

摘要： A bacterium strain B26 capable of producing(R)-α-hydroxyphenylacetic acid [(R)-HPA](yield, 47.5%;enantiomeric excess, 99.1%) from phenylglyoxylic acid(PGA) with high optical purity was isolated and identified as Bacillus sp.B26 by 16S rDNA(ribosomal DNA) sequencing.Phylogenic analysis showed that the strain was most similar to Bacillus sp.enrichment culture clone SYW5(FJ601635.1) and Bacillus cereus strain FM-4(EU794727.1).Efforts were made to further improve HPA-production and PGA-tolerance by UV irradiation and UV-LiCl cooperative mutagenesis.Among viable mutants, B.sp.UV-38 and B.sp.ULi-11 exhibited better productivities than the wild type.Comparisons of HPA production and time course among wild strain and two mutants showed that B.sp.ULi-11 was more competent than B.sp.UV-38.HPA production was increased by 39.1% with B.sp.ULi-11(yield, 65.4%) compared to that with B.sp.B26(yield, 47.0%) when cultured in fermentation broth(pH 7.2) at 32℃ with an agitation speed of 180 r·min^-1 and PGA final concentration of 15 mmol·L^-1 for 25 h.

关键词: (R)-α-hydroxyphenylacetic acid, biotransformation, phylogenetic analysis, Bacillus, phenylglyoxylic acid, UV mutagenesis

CHEN Bingmei, XU Xiaoping, HOU Zhiguo, LI Zhongqin, RUAN Wenbing. Identification and Mutagenesis of a New Isolated Strain Bacillus sp. B26 for Producing(R)-α-Hydroxyphenylacetic Acid [J]. , 2011, 19(4): 636-643.

陈秉梅, 许小平, 侯志国, 李忠琴, 阮文兵. Identification and Mutagenesis of a New Isolated Strain Bacillus sp. B26 for Producing(R)-α-Hydroxyphenylacetic Acid [J]. , 2011, 19(4): 636-643.

References

1 Whitesell, J.K., Reynolds, D., “Resolution of chiral alcohols with mandelic acid”, J. Org. Chem., 48 (20), 3548-3551 (1983).
2 Furlenmeier, A., Quitt, P., Vogler, K., Lanz, P., “6-Acyl derivatives of aminopenicillanic acid”, U.S. Pat., 3957758 (1976).
3 Mills, J., Schmiegel, K.K., Shaw, W.N., “Phenethanolamines, compositions containing the same and method for effecting weight control”, U.S. Pat., 4391826 (1989).
4 Yadav, G.D., Sivakumar, P., “Enzyme-catalyzed optical resolution of mandelic acid via RS-(±) methyl mandelate in non-aqueous media”, Biochem. Eng. J., 19 (2), 101-107 (2004).
5 Ebert, C., Ferluga, G., Gardossi, L., Gianferrara, T., Linda, P., “Improved lipase-mediated resolution of mandelic acid esters by multivariate investigation of experimental factors”, Tetrahedron:Asymmetry, 3 (7), 903-912 (1992).
6 Palomo, J.M., Fernandez-Lorente, G., Mateo, C., Ortiz, C., Fernandez-Lafuente, R., Guisan, J.M., “Modulation of the enantioselectivity of lipases via controlled immobilization and medium engineering:Hydrolytic resolution of mandelic acid esters”, Enzyme Microb. Technol., 31 (6), 775-783 (2002).
7 Huang, H.R., Xu, J.H., Xu, Y., Pan, J., Liu, X., “Preparation of (S)-mandelic acids by enantioselective degradation of racemates with a new isolate Pseudomonas putida ECU1009”, Tetrahedron:Asymmetry, 16 (12), 2113-2117 (2005).
8 Evans, D.A., Morrissey, M.M., Dorow, R.L., “Asymmetric oxygenation of chiral imide enolates. A general approach to the synthesis of enantiomerically pure α-hydroxy carboxylic acid synthons”, J. Am. Chem. Soc., 107 (14), 4346-4348 (1985).
9 Endo, Takakazu, Tamura, Koji, “Process for producing R(-)-mandelic acid or a derivative thereof from a mandelonitrile using rhodococcus”, U.S. Pat., 5296373 (1994).
10 Mori, T., Furui, M., Nakamichi, K., Takahashi, E., “Process for producing D-mandelic acid from benzoylformic acid”, U.S. Pat., 5441888 (1995).
11 Patel, R.N., “Biocatalytic synthesis of intermediates for the synthesis of chiral drug substances”, Curr. Opin. Biotechnol., 12 (6), 587-604 (2001).
12 Choi, W.J., Lee, K.Y., Kang, S.H., Lee, S.B., “Biocatalytic enantioconvergent separation of racemic mandelic acid”, Sep. Puri. Technol., 53 (2), 178-182 (2007)
13 Hirakawa, H., Kamiya, N., Yata, T., Nagamune, T., “Regioselective reduction of a steroid in a reversed micellar system with enzymatic NADH-regeneration”, J. Biochem. Eng., 16 (1), 35-40 (2003).
14 Delecouls-Servat, K., Basseguy, R., Bergel, A., “Membrane electrochemical reactor (MER):Application to NADH regeneration for ADH-catalysed synthesis”, Chem. Eng. Sci., 57 (21), 4633-4642 (2002).
15 Stewart, J.D., “Organic transformations catalyzed by engineered yeast cells and related systems”, Curr. Opin. Biotechnol., 11 (4), 363-368 (2000).
16 Sambrook, J., Fritsch, E.F., Maniatis, T., Molecular cloning. A laboratory manual. Cold Spring Harbor, Cold Spring Harbor Laboratory Press, New York, 127-128 (1989).
17 Tamura, K., Dudley, J., Nei, M., Kumar, S., “MEGA4:Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0”, Mol. Biol Evol, 24, 1596-1599 (2007).
18 Saitou, N., Nei, M., “The neighbor-joining method:A new method for reconstructing phylogenetic trees”, Mol. Biol Evol., 4, 406-425 (1987).
19 Felsenstein, J., “Confidence limits on phylogenies:An approach using the bootstrap”, Evolution, 39, 783-791 (1985).
20 Kannappan, P.G., Shreela, M., Joanna, A., Karuppan, M., “Bacillus sp. mutant for improved biodegradation of Congo red:Random mutagenesis approach”, Bioresour. Technol., 100 (24), 6295-6300 (2009).
21 Drancourt, M., Bollet, C., Carlioz, R., “16S ribosomal DNA sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates”, J. Clin. Microbiol., 38 (10), 3623-3630 (2000).
22 Mahmood, Q., Hu, B.L., Cai, J., Zheng, P., Azim, M.R., “Isolation of Ochrobactrum sp. QZ2 from sulfide and nitrite treatment system”, J. Hazar. Mater., 165, 558-565 (2009)
23 Xiao, M.T., Huang, Y.Y., Meng, C., Guo, Y.H., “Kinetics of asymmetric reduction of phenylglyoxylic acid to R-(-)-mandelic acid by Saccharomyces Cerevisiae FDllb”, Chin. J. Chem. Eng., 14 (1), 73-80 (2006).

[1]	Yan Zhang, Bekir Engin Eser, Peter Kristensen, Zheng Guo. Fatty acid hydratase for value-added biotransformation: A review [J]. Chinese Journal of Chemical Engineering, 2020, 28(8): 2051-2063.
[2]	Jintao Guan, Chaofei Han, Yixin Guan, Songhong Zhang, Junxian Yun, Shanjing Yao. Optimizational production of phenyllactic acid by a Lactobacillus buchneri strain via uniform design with overlay sampling methodology [J]. Chin.J.Chem.Eng., 2019, 27(2): 418-425.

Identification and Mutagenesis of a New Isolated Strain Bacillus sp. B26 for Producing(R)-α-Hydroxyphenylacetic Acid

Identification and Mutagenesis of a New Isolated Strain Bacillus sp. B26 for Producing(R)-α-Hydroxyphenylacetic Acid

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 2

Recommended Articles

Metrics

Comments