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High Yield Production of a Soluble Bifidobacterial β-Galactosidase (BbgIV) in E. coli DH5α with Improved Catalytic Efficiency for the Synthesis of Prebiotic Galactooligosaccharides

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논문

High Yield Production of a Soluble Bifidobacterial β-Galactosidase (BbgIV) in E. coli DH5α with Improved Catalytic Efficiency for the Synthesis of Prebiotic Galactooligosaccharides

학술지

Journal of agricultural and food chemistry

저자명

Osman, Ali; Tzortzis, George; Rastall, Robert A.; Charalampopoulos, Dimitris

초록

<P>The bifidobacterial &beta;-galactosidase (BbgIV) was produced in <I>E. coli</I> DH5&alpha; at 37 and 30 &deg;C in a 5 L bioreactor under varied conditions of dissolved oxygen (dO2) and pH. The yield of soluble BbgIV was significantly (<I>P</I> < 0.05) increased once the dO2 dropped to 0&ndash;2% and remained at such low values during the exponential phase. Limited dO2 significantly (<I>P</I> < 0.05) increased the plasmid copy number and decreased the cells growth rate. Consequently, the BbgIV yield increased to its maximum (71&ndash;75 mg per g dry cell weight), which represented 20&ndash;25% of the total soluble proteins in the cells. In addition, the specific activity and catalytic efficiency of BbgIV were significantly (<I>P</I> < 0.05) enhanced under limited dO2 conditions. This was concomitant with a change in the enzyme secondary structure, suggesting a link between the enzyme structure and function. The knowledge generated from this work is very important for producing BbgIV as a biocatalyst for the development of a cost-effective process for the synthesis of prebiotic galactooligosaccharides from lactose.</P><P><B>Graphic Abstract</B><BR><IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jafcau/2013/jafcau.2013.61.issue-9/jf304792g/production/images/medium/jf-2012-04792g_0005.gif'></P>

발행연도

2013

발행기관

American Chemical Society

ISSN

0021-8561

ISSN

1520-5118

61

9

페이지

pp.2213-2223

주제어

β-galactosidase; Bifidobacterium; protein expression; dissolved oxygen; prebiotic galactooligosaccharides; specific activity

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1 2023-12-11

논문; 2013-12-31

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