Abstract：
:Cellobiose phosphorylase (CBP) cleaves cellobiose-abundant in plant biomass-to glucose and glucose 1-phosphate. However, the pentose sugar xylose, also abundant in plant biomass, acts as a mixed-inhibitor and a substrate for the reverse reaction, limiting the industrial potential of CBP. Preventing xylose, which lacks only a single hydroxymethyl group relative to glucose, from binding to the CBP active site poses a spatial challenge for protein engineering, since simple steric occlusion cannot be used to block xylose binding without also preventing glucose binding. Using CRISPR-based chromosomal library selection, we identified a distal mutation in CBP, Y47H, responsible for improved cellobiose consumption in the presence of xylose. In silico analysis suggests this mutation may alter the conformation of the cellobiose phosphorylase dimer complex to reduce xylose binding to the active site. These results may aid in engineering carbohydrate phosphorylases for improved specificity in biofuel production, and also in the production of industrially important oligosaccharides.
journal_name
ACS Synth Bioljournal_title
ACS synthetic biologyauthors
Chomvong K,Lin E,Blaisse M,Gillespie AE,Cate JHdoi
10.1021/acssynbio.6b00211subject
Has Abstractpub_date
2017-02-17 00:00:00pages
206-210issue
2issn
2161-5063journal_volume
6pub_type
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journal_title：ACS synthetic biology
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journal_title：ACS synthetic biology
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journal_title：ACS synthetic biology
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journal_title：ACS synthetic biology
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journal_title：ACS synthetic biology
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doi：10.1021/acssynbio.9b00192
更新日期：2019-07-19 00:00:00
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doi：10.1021/acssynbio.0c00128
更新日期：2020-07-17 00:00:00
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journal_title：ACS synthetic biology
pub_type： 杂志文章,评审
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更新日期：2014-10-17 00:00:00
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journal_title：ACS synthetic biology
pub_type： 杂志文章
doi：10.1021/acssynbio.6b00388
更新日期：2017-07-21 00:00:00
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journal_title：ACS synthetic biology
pub_type： 杂志文章
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更新日期：2018-02-16 00:00:00
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journal_title：ACS synthetic biology
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pub_type： 杂志文章
doi：10.1021/acssynbio.9b00030
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pub_type： 杂志文章
doi：10.1021/acssynbio.7b00253
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