Abstract:
:We generated a bifunctional enzyme chimera containing the xylanase and lichenase coupled with SpyTag between them. Meanwhile, we generated another chimera containing SpyCatcher and elastin-like polypeptides (ELPs). As ELPs could bond to the xylanase-lichenase chimera through SpyTag/SpyCatcher spontaneous reaction in mild condition, which would lead to the formation of a 3-arm star multifunctional chimera. We purified the xylanase-lichenase by the non-chromatographic purification tag of ELPs. Interestingly, 57.5% of the xylanase and 47.2% of the lichenase in chimera self-assembled into insoluble active particles during the process of purification, which could serve as immobilized bifunctional enzymes. Notably, the immobilized chimera xylanase-lichenase showed a remarkable stability even after 10 reaction cycles, which retained around 56% (lichenase) and 44% (xylanase) of their initial activities, respectively. Moreover, the enhanced thermostability of the immobilized enzymes was also achieved. After incubating at 60 °C for 60 min, the residual activity of the immobilized lichenase was 35%, while the free one was only 24%. Unexpectedly, the free xylanase almost lost its activity when incubated at 55 °C for 60 min, whereas the immobilized xylanase retained 10% of its activity. However, the catalytic efficiency (kcat/Km) of the free xylanase was 1.7-fold higher than the immobilized one, while the free lichenase was 1.1-fold higher than the immobilized one. This is among the first known reports that two enzymes are purified and immobilized in one-step. This novel strategy is easy to scale up and may meet the demands of biofuel industry. It would have great potentials in other biotechnological fields, such as the multifunctional biomaterials systems.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Lin Y,Jin W,Wang J,Cai Z,Wu S,Zhang Gdoi
10.1016/j.enzmictec.2018.04.007subject
Has Abstractpub_date
2018-08-01 00:00:00pages
29-36eissn
0141-0229issn
1879-0909pii
S0141-0229(18)30089-9journal_volume
115pub_type
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.06.014
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.12.004
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.06.002
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.06.015
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
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