Abstract:
BACKGROUND:The Bacillus subtilis spore has long been used to display antigens and enzymes. Spore display can be accomplished by a recombinant and a non-recombinant approach, with the latter proved more efficient than the recombinant one. We used the non-recombinant approach to independently adsorb two thermophilic enzymes, GH10-XA, an endo-1,4-β-xylanase (EC 3.2.1.8) from Alicyclobacillus acidocaldarius, and GH3-XT, a β-xylosidase (EC 3.2.1.37) from Thermotoga thermarum. These enzymes catalyze, respectively, the endohydrolysis of (1-4)-β-D-xylosidic linkages of xylans and the hydrolysis of (1-4)-β-D-xylans to remove successive D-xylose residues from the non-reducing termini. RESULTS:We report that both purified enzymes were independently adsorbed on purified spores of B. subtilis. The adsorption was tight and both enzymes retained part of their specific activity. When spores displaying either GH10-XA or GH3-XT were mixed together, xylan was hydrolysed more efficiently than by a mixture of the two free, not spore-adsorbed, enzymes. The high total activity of the spore-bound enzymes is most likely due to a stabilization of the enzymes that, upon adsorption on the spore, remained active at the reaction conditions for longer than the free enzymes. Spore-adsorbed enzymes, collected after the two-step reaction and incubated with fresh substrate, were still active and able to continue xylan degradation. The recycling of the mixed spore-bound enzymes allowed a strong increase of xylan degradation. CONCLUSION:Our results indicate that the two-step degradation of xylans can be accomplished by mixing spores displaying either one of two required enzymes. The two-step process occurs more efficiently than with the two un-adsorbed, free enzymes and adsorbed spores can be reused for at least one other reaction round. The efficiency of the process, the reusability of the adsorbed enzymes, and the well documented robustness of spores of B. subtilis indicate the spore as a suitable platform to display enzymes for single as well as multi-step reactions.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Mattossovich R,Iacono R,Cangiano G,Cobucci-Ponzano B,Isticato R,Moracci M,Ricca Edoi
10.1186/s12934-017-0833-3subject
Has Abstractpub_date
2017-11-28 00:00:00pages
218issue
1issn
1475-2859pii
10.1186/s12934-017-0833-3journal_volume
16pub_type
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 社论
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journal_title:Microbial cell factories
pub_type: 社论
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abstract:BACKGROUND:L-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In Escherichia coli, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzymes in the shikimate pathway and L-tryptophan branch pathway, while L-serine and phosphoribosylpyr...
journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0410-1
更新日期:2016-01-14 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-13-11
更新日期:2014-01-17 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-7
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2021-01-19 00:00:00
abstract:BACKGROUND:Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study...
journal_title:Microbial cell factories
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更新日期:2010-06-17 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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abstract:BACKGROUND:The methylotrophic yeast Pichia pastoris is a common host for the production of recombinant proteins. However, hypermannosylation hinders the use of recombinant proteins from yeast in most biopharmaceutical applications. Glyco-engineered yeast strains produce more homogeneously glycosylated proteins, but can...
journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
pub_type: 杂志文章
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abstract:BACKGROUND:Although the citric acid fermentation by Aspergillus niger is one of the most important industrial microbial processes and various aspects of the fermentation appear in a very large number of publications since the 1950s, the effect of the spore inoculum level on fungal morphology is a rather neglected area....
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-5-3
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