Two-step biosynthesis of d-allulose via a multienzyme cascade for the bioconversion of fruit juices

Li, Chao; Lei, Li; Feng, Zhiyuan; Guan, Liming; Lu, Fuping; Qin, Hao

doi:10.1016/j.foodchem.2021.129746

Cited by 39 publications

(23 citation statements)

References 34 publications

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“…Moreover, the SfDAE WT and the M15S/P40N/S209N variant were immobilized on the epoxy support ES-103B as described in our previous report . The storage stabilities of the immobilized SfDAE WT and M15S/P40N/S209N variant were analyzed by incubating them at 4 °C for 28 days, respectively.…”

Section: Materials and Methodsmentioning

confidence: 99%

Continuous Spectrophotometric Assay for High-Throughput Screening of Predominant d-Allulose 3-Epimerases

Zhang

Wei

et al. 2021

J. Agric. Food Chem.

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d-Allulose is an attractive noncaloric sugar substitute with numerous health benefits, which can be biosynthesized by d-allulose 3-epimerases (DAEases). However, enzyme instability under harsh industrial reaction conditions hampered its practical applications. Here, we developed a continuous spectrophotometric assay (CSA) for the efficient analysis of d-allulose in a mixture. Furthermore, a high-throughput screening strategy for DAEases was developed using CSA by coupling DAEase with a NADH-dependent ribitol dehydrogenase, enabling high-throughput screening of DAEase variants with desired properties. The variant M15S/P40N/S209N exhibited a half-life of 22 h at 60 °C and an 8.7 °C increase of the T 50 60 value, with a 1.2-fold increase of activity. Structural modeling and molecular dynamics simulations indicated that the improvement of thermostability and activity was due to some new hydrogen bonds between chains at the dimer interface and between the residue and the substrate d-fructose. This work offers a robust tool and theoretical basis for the improvement of DAEases, which will benefit the enzymatic biosynthesis of d-allulose and promote its industrial application.

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Section: Materials and Methodsmentioning

confidence: 99%

Continuous Spectrophotometric Assay for High-Throughput Screening of Predominant d-Allulose 3-Epimerases

Zhang

Wei

et al. 2021

J. Agric. Food Chem.

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“…d -Allulose 3-epimerase (DAEase, EC 5.1.3.30) is one of the most important members of the KEase family and has a pivotal role in the biosynthesis of d -allulose due to the high catalytic activity toward d -fructose relative to other KEase family members. To date, about 20 microorganisms have been reported as the sources of DAEase, and nearly half were identified in the last 3 years, such as Paenibacillus senegalensis, Staphylococcus aureus, Rhodopirellula baltica, DaeM, Thermoclostridium caenicola, Bacillus sp., Pirellula sp., Novibacillus thermophilus, Halanaerobium congolense, and Sinorhizobium fredii …”

Section: Introductionmentioning

confidence: 99%

Computer-Aided Targeted Mutagenesis of Thermoclostridium caenicola d-Allulose 3-Epimerase for Improved Thermostability

Chen

et al. 2022

J. Agric. Food Chem.

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D-Allulose 3-epimerase (DAEase) is a key enzyme in D-allulose bioproduction. DAEase from Thermoclostridium caenicola suffers from poor thermostability, hampering its large-scale applications in industry. In this study, mutants A70P, G107P, F155Y, and D162T with increased melting point temperature (T m ) were obtained by targeted mutagenesis based on the calculation of the free energy of folding. The optimal single-point mutant G107P showed 11.08 h, 5, and 5.70 °C increases in the values of halflife (t 1/2 ) at 60 °C, the optimum temperature (T opt ), and T m , respectively. Beneficial mutations were combined by ordered recombination mutagenesis, and the combinational mutant Var3 (G107P/F155Y/D162T/A70P) was generated with ΔT opt of 10 °C and ΔT m of 12.25 °C. Its t 1/2 value at 65 °C was more than 140 times higher than that of the wild-type enzyme. Molecular dynamics simulations and homology modeling analysis indicated that the enhanced overall rigidity, increased hydrogen bonds between subunits, and redistributed surface electrostatic charges might be responsible for the improved thermostability of the mutant Var3.

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“…The most popular strategy for d ‐allulose synthesis (Izumoring strategy) is through the epimerization of d ‐fructose at C‐3 position catalyzed by ketose 3‐epimerase (KEase) 19 . Among members of the KEase family, d ‐allulose/ d ‐psicose 3‐epimerase (DAEase/DPEase) and d ‐tagatose 3‐epimerase (DTEase) are mainly used for d ‐allulose production 20‐27 . Compared with DTEase, DAEase usually exhibits high catalytic efficiency for d ‐allulose synthesis.…”

Section: Introductionmentioning

confidence: 99%

“…19 Among members of the KEase family, D-allulose/D-psicose 3-epimerase (DAEase/DPEase) and D-tagatose 3-epimerase (DTEase) are mainly used for D-allulose production. [20][21][22][23][24][25][26][27] Compared with DTEase, DAEase usually exhibits high catalytic efficiency for D-allulose synthesis. Nevertheless, the conversion rate of D-allulose by KEase is unsatisfactory due to the limitation of thermodynamic equilibrium.…”

Section: Introductionmentioning

confidence: 99%

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

Feng

Zhou

et al. 2022

J Sci Food Agric

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BACKGROUND Rare sugars have become promising ‘sugar alternatives’ because of their low calories and unique physiological functions. Among the family of rare sugars, d‐allulose is one of the sugars attracting interest. Ketose 3‐epimerases (KEase), including d‐tagatose 3‐epimerase (DTEase) and d‐allulose 3‐epimerase (DAEase), are mainly used for d‐allulose production. RESULTS In this study, a putative xylose isomerase from Caballeronia insecticola was characterized and identified as a novel DAEase. Caballeronia insecticola DAEase displayed prominent enzymatic properties, and 150 g L−1 d‐allulose was produced from 500 g L−1 d‐fructose in 45 min with a conversion rate of 30% and high productivity of 200 g L−1 h−1. Furthermore, DAEase was employed in a phosphorylation–dephosphorylation cascade reaction, which significantly increased the conversion rate of d‐allulose. Under optimized conditions, the conversion rate of d‐allulose was approximately 100% when the concentration of d‐fructose was 50 mmol L−1. CONCLUSION This research described a very beneficial and facile approach for d‐allulose production based on C. insecticola DAEase. © 2022 Society of Chemical Industry.

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Two-step biosynthesis of d-allulose via a multienzyme cascade for the bioconversion of fruit juices

Cited by 39 publications

References 34 publications

Continuous Spectrophotometric Assay for High-Throughput Screening of Predominant d-Allulose 3-Epimerases

Continuous Spectrophotometric Assay for High-Throughput Screening of Predominant d-Allulose 3-Epimerases

Computer-Aided Targeted Mutagenesis of Thermoclostridium caenicola d-Allulose 3-Epimerase for Improved Thermostability

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

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