Preparation of recombinant L-glutamate oxidase immobilized cellsLI Li1,WANG Ya-ping1,RAO Ben2,MA Li-xin1

doi:10.14088/j.cnki.issn0439-8114.2019.17.035

HUBEI AGRICULTURAL SCIENCES ›› 2019, Vol. 58 ›› Issue (17): 132-135.doi: 10.14088/j.cnki.issn0439-8114.2019.17.035

• Biological Engineering • Previous Articles Next Articles

Preparation of recombinant L-glutamate oxidase immobilized cellsLI Li¹,WANG Ya-ping¹,RAO Ben²,MA Li-xin¹

Abstract： In order to improve the substrate catalytic tolerance of recombinant E. coli（pET-23a-lgox） expressing L-glutamate oxidase（LGOX）, this study attempted to study the immobilization of recombinant bacteria. The results showed that the best immobilization conditions were 2.5% sodium alginate, 13 g/L CaCl₂ solution, pH 6.5, and the optimal volume ratio of engineering bacteria（OD_{600 nm}=14） to sodium alginate was 1∶;
40. At this time, the mechanical strength of the stationary engineering bacteria was the best. Through the comparison of enzymatic properties, it was found that the immobilized cells were more tolerant than free cells in the substrate catalysis process. And under the same conditions, their temperature tolerance, pH tolerance, and the use of frequency were higher than free cells.

Received:2019-02-21 Published:2019-11-14

Abstract

CLC Number:

Q814.2

（1.Hubei Key Laboratory of Industrial Biotechnology, Hubei University, Wuhan430062, China;2.National Biopesticide Engineering Research Center, Hubei Academy of Agricultural Sciences, Wuhan430064, China）. Preparation of recombinant L-glutamate oxidase immobilized cellsLI Li¹,WANG Ya-ping¹,RAO Ben²,MA Li-xin¹[J]. HUBEI AGRICULTURAL SCIENCES, 2019, 58(17): 132-135.

References

[1] KYEONG J S,YEOM S H.Preparation of immobilized whole cell biocatalyst and biodiesel production using a packed-bed bioreactor[J].Bioprocess Biosyst Eng,2014,37(11):2189-2198.
[2] CHEN J P,LIN G H.Optimization of biodiesel production catalyzed by fungus cells immobilized in fibrous supports[J].Appl Biochem Biotechnol,2010,161(1-8):181-194.
[3] 孙婷,杜伟,刘德华,等.固定化全细胞催化可再生油脂合成生物柴油的稳定性[J].生物工程学报,2009,25(9):1379-1385.
[4] BAN K,KAIEDA M.Whole cell biocatalyst for biodiesel fuel production utilizing Rhizopus oryzae cells immobilized within biomass support particles[J].Biochem Eng J,2001,8(1):39-43.
[5] 刘秀伟,司芳.酶固定化研究进展[J].化工技术经济,2003, 21(4):12-16.
[6] 许松伟,张杰,杨占平,等.载体对包埋酶微环境的影响分析[J].化学进展,2008,20(1):163-170.
[7] 蒋挺大. 甲壳素和壳聚糖作为酶和细胞的固定化载体的研究进展[J].工业微生物,1998,28(3):32-37.
[8] ROGER A,SHELDON. Enzyme immobilization:The quest for optimum performance[J].Reviews,2007,10(1002):1289-1307.
[9] SERIKOV V S,LIASHEV I D.The influence of melatonin on the content of lipid peroxidation metabolites and antioxidant enzyme activity in the liver of rats after immobilization[J].Ross Fiziol Zh Im I M Sechenova,2014,100(4):458-464.
[10] ARIMA J,TAMURA T,KUSAKABE H,et al.Recombinant expression,biochemical characterization and stabilizationthrough proteolysis of an l-glutamate oxidase from Streptomyces sp. X-119-6[J].J Biochem,2003,134:805-812.
[11] WACHIRATIANCHAI S,BHUMIRATANA A,UDOMSOPAGIT S.Isolation,purification and characterization of l-glutamate oxidase from Streptomyces sp. 18G[M].Electron J Biotech,2004,7(3):9-10.
[12] 李青山,李晓波,李友荣,等.用静息细胞培养法研究L-谷氨酸氧化酶的生物合成[J].华东理工大学学报,1994,20(2):168-172.
[13] YAPING W, BEN R,HONG Y,et al.High-level expression of l-glutamate oxidase in Pichia pastoris using multi-copy expression strains and high cell density cultivation[J].Protein expression and purification,2017,129:108-114.
[14] 张飞,刘寅,魏涛,等.缬氨酸转氨酶固定化细胞的制备[J].中国调味品,2014,39(8):9-15
[15] 韩振为,言海燕.海藻酸钠-硅藻土包埋石油脱硫菌Rhodococcus sp. H-412[J].化学工业与工程,2006,23(6):520-522.

Preparation of recombinant L-glutamate oxidase immobilized cellsLI Li¹,WANG Ya-ping¹,RAO Ben²,MA Li-xin¹

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 0

Recommended Articles

Metrics

Comments