Efficient biosynthesis of D-allulose in Bacillus subtilis through D-psicose 3-epimerase translation modification

Zhao, Jingyi; Wei, Hongbei; Chen, Jing; Li, Lihong; Li, Kai; Liu, Jidong

doi:10.1016/j.ijbiomac.2021.07.093

Cited by 14 publications

(10 citation statements)

References 38 publications

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“…Moreover, the activity was significantly reduced at pH values below 8.0 and decreased to 20% of the maximum activity at pH 6.0. Similar to other reported DAEase proteins, 21,27 C. insecticola DAEase is a metaldependent enzyme, and the optimal metal ion was Mn 2+ (Fig. 5C).…”

Section: Optimization Of Reaction Conditions and D-allulose Productio...supporting

confidence: 89%

“…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%

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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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Section: Optimization Of Reaction Conditions and D-allulose Productio...supporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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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“…To date, several endogenous constitutive promoters are known in B. subtilis , such as P HpaII , P 43 , P srfA , and P spovG [ 20 − 22 ]. But most researchers are accustomed to using the same promoter to express different genes, which results in the low production of target protein expression [ 23 , 24 ]. Therefore, auto-inducible promoter engineering is necessary to develop various cassettes to enhance protein yields and improve specific enzyme activity.…”

Section: Introductionmentioning

confidence: 99%

Efficient D-allulose synthesis under acidic conditions by auto-inducing expression of the tandem D-allulose 3-epimerase genes in Bacillus subtilis

Wei

Zhang

et al. 2022

Microb Cell Fact

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Background D-allulose, a hexulose monosaccharide with low calorie content and high sweetness, is commonly used as a functional sugar in food and nutrition. However, enzyme preparation of D-allulose from D-frutose was severely hindered by the non-enzymatic browning under alkaline and high-temperature, and the unnecessary by-products further increased the difficulties in separation and extraction for industrial applications. Here, to address the above issue during the production process, a tandem D-allulose 3-epimerase (DPEases) isomerase synergistic expression strategy and an auto-inducible promoter engineering were levered in Bacillus subtilis 168 (Bs168) for efficient synthesis of D-allulose under the acidic conditions without browning. Results First, based on the dicistron expression system, two DPEases with complementary functional characteristics from Dorea sp. CAG:317 (DSdpe) and Clostridium cellulolyticum H10 (RCdpe) were expressed in tandem under the promoter HpaII in one cell. A better potential strain Bs168/pMA5-DSdpe-RCdpe increases enzyme activity to 18.9 U/mL at acidic conditions (pH 6.5), much higher than 17.2 and 16.7 U/mL of Bs168/pMA5-DSdpe and Bs168/pMA5-RCdpe, respectively. Subsequently, six recombinant strains based on four constitutive promoters were constructed in variable expression cassettes for improving the expression level of protein. Among those engineered strains, Bs168/pMA5-PspoVG-DSdpe-PsrfA-RCdpe exhibited the highest enzyme activity with 480.1 U/mL on fed-batch fermentation process in a 5 L fermenter at pH 6.5, about 2.1-times higher than the 228.5 U/mL of flask fermentation. Finally, the maximum yield of D-allulose reached as high as 163.5 g/L at the fructose concentration (50% w/v) by whole-cell biocatalyst. Conclusion In this work, the engineered recombinant strain Bs168/pMA5-PspoVG-DSdpe-PsrfA-RCdpe was demonstrated as an effective microbial cell factory for the high-efficient synthesis of D-allulose without browning under acidic conditions. Based on the perspectives from this research, this strategy presented here also made it possible to meet the requirements of the industrial hyper-production of other rare sugars under more acidic conditions in theory.

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“…D -Allulose is a low-energy sweet and is regarded as a potential substitute for sucrose as it has 70% of the relative sweetness but only 0.3% of the energy of sucrose ( Zhang et al, 2015 ). More importantly, D -allulose has many important physiological functions, for example, blood glucose suppressive effect ( EdyLiani et al, 2020 ), body fat accumulation inhibitive effect ( Kim et al, 2017 ), reactive oxygen species scavenging effect ( Li et al, 2018 ), and neuroprotective effect ( Zhao et al, 2021 ). In addition, it has good properties for food industry applications, such as improving the gelling behavior and producing good flavor ( Zhang et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

D-Allulose (D-Psicose) Biotransformation From Allitol by a Newly Found NAD(P)-Dependent Alcohol Dehydrogenase From Gluconobacter frateurii NBRC 3264 and the Enzyme Characterization

Wen¹,

Lin²,

Ning³

et al. 2022

Front. Microbiol.

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The NAD(P)-dependent alcohol dehydrogenase (ADH) gene was cloned from Gluconobacter frateurii NBRC 3264 and expressed in Escherichia coli BL21 star (DE3). The expressed enzyme was purified and the characteristics were investigated. The results showed that this ADH can convert allitol into D-allulose (D-psicose), which is the first reported enzyme with this catalytic ability. The optimum temperature and pH of this enzyme were 50°C and pH 7.0, respectively, and the enzyme showed a maximal activity in the presence of Co2+. At 1 mM Co2+ and allitol concentrations of 50, 150, and 250 mM, the D-allulose yields of 97, 56, and 38%, respectively, were obtained after reaction for 4 h under optimal conditions, which were much higher than that obtained by using the epimerase method of about 30%.

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Efficient biosynthesis of D-allulose in Bacillus subtilis through D-psicose 3-epimerase translation modification

Cited by 14 publications

References 38 publications

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

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

Efficient D-allulose synthesis under acidic conditions by auto-inducing expression of the tandem D-allulose 3-epimerase genes in Bacillus subtilis

D-Allulose (D-Psicose) Biotransformation From Allitol by a Newly Found NAD(P)-Dependent Alcohol Dehydrogenase From Gluconobacter frateurii NBRC 3264 and the Enzyme Characterization

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