Efficient enzymatic synthesis of <scp>d</scp>‐allulose using a novel <scp>d</scp>‐allulose‐3‐epimerase from <i>Caballeronia insecticola</i>

Li, Zijie; Feng, Linxue; Zhou, Chen; Fei, Kangqing; Xu, Haitao; Gao, Xiao‐Dong

doi:10.1002/jsfa.12147

Cited by 13 publications

(5 citation statements)

References 33 publications

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“…In the search for new DAEase, the amino acid sequence of Pseudomonas cichorii DTEase was used as a template for the NCBI BLAST tool. Li et al [ 16 ] discovered a putative xylose isomerase (XI) from C. insecticola (GenBank Accession: BAN26336.1) after various in silico analyses. The resultant expressed CiDAEase had a slightly better conversion rate of up to 31% at the optimal temperature of 65 °C with 500 g/L of D -fructose after 45 min, as compared to ApDAEase.…”

Section: Characterization Of Newly Identified Daeasesmentioning

confidence: 99%

“…While the addition of Mg 2+ also improved activity, the optimal metal ion for CiDAEase was Mn 2+ . When used in a multienzyme cascade reaction, a conversion rate of more than 80% was achieved in 5 h [ 16 ].…”

Section: Characterization Of Newly Identified Daeasesmentioning

confidence: 99%

“…In this review, we discussed how new epimerases can be discovered [ 15 , 16 , 17 ], as well as methods to improve their stability and activity via protein engineering [ 21 , 22 , 24 , 25 , 28 ], the application of B. subtilis [ 11 , 25 , 29 ], and the potential of immobilization techniques to enhance performance [ 34 , 35 , 36 , 37 ]. These advancements have the potential to significantly reduce production time and costs associated with D -allulose biosynthesis.…”

Section: Summary and Future Visionmentioning

confidence: 99%

“…In the last two years, there have been at least three new notable identified DAEase that were reported and characterized (Table 1). The DAEase from Arthrobacter psychrolactophilus B7 (ApDAEase) [15], Caballeronia insecticola (CiDAEase) [16], and DPEase from Iocasia fonsfrigidae SP3-1 (IfDPEase) [17] were expressed in Escherichia coli BL21(DE3) and subsequently purified and characterized. There are two general methods that have been established, namely, the use of immobilized enzymes and that of whole-cell biotransformation.…”

Section: Characterization Of Newly Identified Daeasesmentioning

confidence: 99%

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The Engineering, Expression, and Immobilization of Epimerases for D-allulose Production

Tan

Chen

et al. 2023

IJMS

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The rare sugar D-allulose is a potential replacement for sucrose with a wide range of health benefits. Conventional production involves the employment of the Izumoring strategy, which utilises D-allulose 3-epimerase (DAEase) or D-psicose 3-epimerase (DPEase) to convert D-fructose into D-allulose. Additionally, the process can also utilise D-tagatose 3-epimerase (DTEase). However, the process is not efficient due to the poor thermotolerance of the enzymes and low conversion rates between the sugars. This review describes three newly identified DAEases that possess desirable properties for the industrial-scale manufacturing of D-allulose. Other methods used to enhance process efficiency include the engineering of DAEases for improved thermotolerance or acid resistance, the utilization of Bacillus subtilis for the biosynthesis of D-allulose, and the immobilization of DAEases to enhance its activity, half-life, and stability. All these research advancements improve the yield of D-allulose, hence closing the gap between the small-scale production and industrial-scale manufacturing of D-allulose.

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Section: Characterization Of Newly Identified Daeasesmentioning

confidence: 99%

Section: Characterization Of Newly Identified Daeasesmentioning

confidence: 99%

Section: Summary and Future Visionmentioning

confidence: 99%

Section: Characterization Of Newly Identified Daeasesmentioning

confidence: 99%

See 2 more Smart Citations

The Engineering, Expression, and Immobilization of Epimerases for D-allulose Production

Tan

Chen

et al. 2023

IJMS

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“…Meanwhile, the Izumoring strategy based on D-allulose 3-epimerase (DAEase) plays a crucial role in D-allulose production from D-fructose [ 12 ]. Subsequently, a series of DAEases have been discovered in many microorganisms [ 13 , 14 , 15 , 16 , 17 , 18 ]. Thus, D-allulose has gradually become one of the most accessible rare sugars.…”

Section: Introductionmentioning

confidence: 99%

Highly Efficient Synthesis of Rare Sugars from Glycerol in Endotoxin-Free ClearColi by Fermentation

Gao

Chen

Nakanishi

et al. 2023

Foods

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Rare sugars possess potential applications as low-calorie sweeteners, especially for anti-obesity and anti-diabetes. In this study, a fermentation biosystem based on the “DHAP-dependent aldolases strategy” was established for D-allulose and D-sorbose production from glycerol in endotoxin-free ClearColi BL21 (DE3). Several engineering strategies were adopted to enhance rare sugar production. Firstly, the combination of different plasmids for aldO, rhaD, and yqaB expression was optimized. Then, the artificially constructed ribosomal binding site (RBS) libraries of aldO, rhaD, and yqaB genes were assembled individually and combinatorially. In addition, a peroxidase was overexpressed to eliminate the damage or toxicity from hydrogen peroxide generated by alditol oxidase (AldO). Finally, stepwise improvements in rare sugar synthesis were elevated to 15.01 g/L with a high yield of 0.75 g/g glycerol in a 3 L fermenter. This research enables the effective production of rare sugars from raw glycerol in high yields.

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Semi‐rational engineering of D‐allulose 3‐epimerase for simultaneously improving the catalytic activity and thermostability based on D‐allulose biosensor

Li,

Hu,

et al. 2024

Biotechnology Journal

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BackgroundD‐Allulose is one of the most well‐known rare sugars widely used in food, cosmetics, and pharmaceutical industries. The most popular method for D‐allulose production is the conversion from D‐fructose catalyzed by D‐allulose 3‐epimerase (DAEase). To address the general problem of low catalytic efficiency and poor thermostability of wild‐type DAEase, D‐allulose biosensor was adopted in this study to develop a convenient and efficient method for high‐throughput screening of DAEase variants.ResultsThe catalytic activity and thermostability of DAEase from Caballeronia insecticola were simultaneously improved by semi‐rational molecular modification. Compared with the wild‐type enzyme, DAEaseS37N/F157Y variant exhibited 14.7% improvement in the catalytic activity and the half‐time value (t1/2) at 65°C increased from 1.60 to 27.56 h by 17.23‐fold. To our delight, the conversion rate of D‐allulose was 33.6% from 500‐g L−1 D‐fructose in 1 h by Bacillus subtilis WB800 whole cells expressing this DAEase variant. Furthermore, the practicability of cell immobilization was evaluated and more than 80% relative activity of the immobilized cells was maintained from the second to seventh cycle.ConclusionAll these results indicated that the DAEaseS37N/F157Y variant would be a potential candidate for the industrial production of D‐allulose.

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Efficient enzymatic synthesis of d‐allulose using a novel d‐allulose‐3‐epimerase from Caballeronia insecticola

Cited by 13 publications

References 33 publications

The Engineering, Expression, and Immobilization of Epimerases for D-allulose Production

The Engineering, Expression, and Immobilization of Epimerases for D-allulose Production

Highly Efficient Synthesis of Rare Sugars from Glycerol in Endotoxin-Free ClearColi by Fermentation

Semi‐rational engineering of D‐allulose 3‐epimerase for simultaneously improving the catalytic activity and thermostability based on D‐allulose biosensor

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