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Evaluation of gene expression and alginate production in response to oxygen transfer in continuous culture of Azotobacter vinelandii

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논문

Evaluation of gene expression and alginate production in response to oxygen transfer in continuous culture of Azotobacter vinelandii

학술지

PloS one

저자명

Dí az-Barrera, Alvaro; Martí nez, Fabiola; Guevara Pezoa, Felipe; Acevedo, Fernando

초록

<P>Alginates are polysaccharides used as food additives and encapsulation agents in biotechnology, and their functional properties depend on its molecular weight. In this study, different steady-states in continuous cultures of <I>A</I>. <I>vinelandii</I> were established to determine the effect of the dilution rate (D) and the agitation rate on alginate production and expression of genes involved in alginate polymerization and depolymerization. Both, the agitation and dilution rates, determined the partitioning of the carbon utilization from sucrose into alginate and CO<SUB>2</SUB> under oxygen-limiting conditions. A low D (0.07 h<SUP>&#x2212;1</SUP>) and 500 rpm resulted in the highest carbon utilization into alginate (25%). Quantitative real-time polymerase chain reaction was used to determine the transcription level of six genes involved in alginate polymerization and depolymerization. In chemostat cultures at 0.07 h<SUP>&#x2212;1</SUP>, the gene expression was affected by changes in the agitation rate. By increasing the agitation rate from 400 to 600 rpm, the <I>algE7</I> gene expression decreased tenfold, whereas <I>alyA1</I>, <I>algL</I> and <I>alyA2</I> gene expression increased between 1.5 and 2.8 times under similar conditions evaluated. Chemostat at 0.07 h<SUP>&#x2212;1</SUP> showed a highest alginate molecular weight (580 kDa) at 500 rpm whereas similar molecular weights (480 kDa) were obtained at 400 and 600 rpm. The highest molecular weight was not explained by changes in the expression of <I>alg8</I> and <I>alg44</I> (genes involved in alginate polymerization). Nonetheless, a different expression pattern observed for lyases could explain the highest alginate molecular weight obtained. Overall, the results suggest that the control of alginate molecular weight in <I>A</I>. <I>vinelandii</I> cells growing in continuous mode is determined by a balance between the gene expression of intracellular and extracellular lyases in response to oxygen availability. These findings better our understanding of the biosynthesis of bacterial alginate and help us progress toward obtain tailor-made alginates.</P>

발행연도

2014

발행기관

Public Library of Science

라이선스

cc-by

ISSN

1932-6203

9

8

페이지

pp.e105993

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논문; 2014-08-27

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