2022
DOI: 10.1021/acs.jafc.2c04800
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Boosting the Heterologous Expression of d-Allulose 3-Epimerase in Bacillus subtilis through Protein Engineering and Catabolite-Responsive Element Box Engineering

Abstract: As a natural sweetener with low calories and various physiological activities, d-allulose has drawn worldwide attention. Currently, d-allulose 3-epimerase (DAEase) is mainly used to catalyze the epimerization of d-fructose to d-allulose. Therefore, it is quite necessary to enhance the food-grade expression of DAEase to meet the surging market demand for d-allulose. In this study, initially, the promising variant H207L/D281G/C289R of Clostridium cellulolyticum H10 DAEase (CcDAEase) was generated by protein engi… Show more

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Cited by 13 publications
(11 citation statements)
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“…As shown in the table, the fermentation activity in fed-batch processes was significantly higher compared to that in shakeflask fermentation. 17 In this study, Clostridium sp. DAEase was screened from nine KEase candidates for expression in B. subtilis WB600.…”
Section: Transcriptional Regulation For Enhancing Clsp-daease Express...mentioning
confidence: 84%
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“…As shown in the table, the fermentation activity in fed-batch processes was significantly higher compared to that in shakeflask fermentation. 17 In this study, Clostridium sp. DAEase was screened from nine KEase candidates for expression in B. subtilis WB600.…”
Section: Transcriptional Regulation For Enhancing Clsp-daease Express...mentioning
confidence: 84%
“…cellulolyticum DAEase variant H207L/D281G/C289R and then applied a CRE box strategy to eliminate the carbon catabolite repression (CCR) effect. Subsequently, the final fermentation activity reached 4971.5 U/mL after the fed-batch fermentation …”
Section: Resultsmentioning
confidence: 99%
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“…d ‐Allulose is a natural sweetener with various physiological activities, and enhancement of its final productivity from low‐cost substrate is a practicable and economic strategy for meeting the increasing market demand of this compound 20,32 . Despite the exploration of the in vitro enzymatic cascade, D‐allulose production has been < 30% as a result of the thermodynamic equilibrium of DAEase.…”
Section: Discussionmentioning
confidence: 99%
“…31 DISCUSSION D-Allulose is a natural sweetener with various physiological activities, and enhancement of its final productivity from low-cost substrate is a practicable and economic strategy for meeting the increasing market demand of this compound. 20,32 Despite the exploration of the in vitro enzymatic cascade, D-allulose production has been < 30% as a result of the thermodynamic equilibrium of DAEase. Construction of cell factory provides an advantageous space for enzymes, preventing unfavorable changes of the protein structure under severe catalytic environment or in non-conventional reaction system.…”
Section: Wileyonlinelibrarycom/jsfamentioning
confidence: 99%