Difference between revisions of "Team:Nankai/Description"

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Poly-γ-glutamic acid (γ-PGA) is an important,  naturally occurring polyamide consisting of D/L-glutamate monomers. Unlike  typical peptide linkages, the amide linkages inγ-PGA are  formed between the α-amino group and the  γ-carboxyl group. γ-PGA exhibits many favorable features such as biodegradable,  water soluble, edible and non-toxic to humans and the environment. Therefore,  it has been widely used in fields of foods, medicines, cosmetics and  agriculture and many unique applications, such as a sustained release material  and drug carrier, curable biological adhesive, biodegradable fibres, and highly  water absorbable hydrogels.<br>
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  Strains capable for producing γ-PGA  are divided into two categories based on their requirement for glutamate acid:  glutamate-dependent strains and glutamate-independent strains.  Glutamate-independent strains are preferable for industrial production because  of their low cost and simplified fermentation process. However, compared with  glutamate-dependent strains, their lower γ-PGA productivity limits their  industrial application.  Therefore,  the construction of a glutamate-independent strain with high γ-PGA yield is  important for industrial applications. <br>
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  <em>Bacillusamyloliquefaciens</em> LL3, isolated from fermented food, is a  glutamate-independent strain, which can produce 3-4 g/L γ-PGA with sucrose as  its carbon source and ammonium sulfate as its nitrogen source. The <em>B. amyloliquefaciens</em> LL3 strain was deposited in the China Center for Type Culture  Collection (CCTCC) with accession number CCTCC M 208109 and its whole genome has been sequenced in 2011. In this study,  we aimed to improve the γ-PGA production based on the <em>B. amyloliquefaciens </em>NK-1 strain (a derivative of LL3 strain with  its endogenous plasmid and <em>upp</em> gene  deleted).<br>
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In order to improve  γ-PGA production, we employed two strategies to fine-tune the synthetic  pathways and balance the metabolism in the glutamate-independent <em>B. amyloliquefaciens </em>NK-1 strain. Firstly,  we constructed a metabolic toggle switch in the NK-1 strain to inhibit the  expression of ODHC (2-oxoglutarate dehydrogenase complex) by adding IPTG in the  stationary stage and distribute the metabolic flux more frequently to be used  for γ-PGA precursor-glutamate synthesis. As scientists had found that the activity of ODHC was rather  low when glutamate was highly produced in a  <em>Corynebacterium&nbsp;glutamicum</em> strain. Second, to balance the increase  of endogenous glutamate production, we optimized the expression level of <em>pgsBCA</em> genes (responsible for γ-PGA synthesis) by replacing its native promoter to  seven different strength of promoters. Through these two strategies, we aimed  to obtain a γ-PGA production improved mutant strain. </p>
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Revision as of 08:26, 4 August 2015

Welcome to our website

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