Food and Fermentation Industries >

2021 , Vol. 47 >Issue 20: 1 - 7

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026871

Cloning, expression and characterization of L-rhamnulose kinase from Shigella flexneri

  • FENG Linxue ,
  • CHEN Zhou ,
  • WANG Yasen ,
  • FENG Kang ,
  • XU Xiangyang ,
  • LI Zijie ,
  • NAKANISHI Hideki ,
  • GAO Xiaodong
Expand
  • 1(School of Bioengineering, Jiangnan University, Wuxi 214122, China)
    2(Zaozhuang Jienuo Biological Enzyme Co.Ltd., Zaozhuang 277100, China)

Received date: 2021-01-21

  Revised date: 2021-03-01

  Online published: 2021-11-18

Fold

Abstract

D-Allulose is a kind of rare sugars with health care function and its production is mainly catalyzed by DPE (D-Psicose 3-epimerase) which can convert D-fructose to D-allulose. However, the conversion rate of this reaction is only about 30%. One method to improve the conversion rate is a phosphorylation-dephosphorylation cascade reaction based on L-rhamnulose kinase (RhaB). Up to now, the research about RhaB has been rarely reported. In this study, a gene encoding RhaB from Shigella flexneri 2a str. 301 was cloned into pET28a and expressed in Escherichia coli BL21(DE3). To enhance the activity of RhaB, we constructed a mutant strain RhaBE437Q. As a consequence, the activity was increased by 10 times and most of the inclusion body proteins became soluble. The purified recombinant RhaBE437Q had the maximum activity at 40 ℃, pH 8.5 and Mn2+, and was identified as a monomer with a molecular weight of 54 kDa. It could only catalyse sugars with the structure of R configuration of C-3. At last, we docked D-allulose into RhaBE437Q and performed a preliminary research on its catalytic mechanism.

Key words： L-rhamnulose kinase; Shigella flexneri 2a str.301; cloning and expressing; enzymatic properties; D-allulose; catalytic mechanism

Cite this article

FENG Linxue , CHEN Zhou , WANG Yasen , FENG Kang , XU Xiangyang , LI Zijie , NAKANISHI Hideki , GAO Xiaodong . Cloning, expression and characterization of L-rhamnulose kinase from Shigella flexneri[J]. Food and Fermentation Industries, 2021 , 47(20) : 1 -7 . DOI: 10.13995/j.cnki.11-1802/ts.026871

References

[1] GRANSTRÖM T B, TAKATA G, TOKUDA M, et al.Izumoring:A novel and complete strategy for bioproduction of rare sugars[J].Journal of Bioscience & Bioengineering, 2004, 97(2):89-94.
[2] 贾东旭, 孙晨奕, 彭晨, 等.D-阿洛酮糖及其合成研究进展[J].食品与发酵工业, 2021, 47(3):211-217.
JIA D X, SUN C Y, PENG C, et al.Advances on D-psicose and its synthesis[J].Food and Fermentation Industries, 2021, 47(3):211-217.
[3] HAN Y, YOON J, CHOI M S.Tracing the anti-inflammatory mechanism/triggers of d-allulose:A profile study of microbiome composition and mRNA expression in diet-induced obese mice[J].Molecular Nutrition & Food Research, 2020, 64(5):1900982.
[4] HAN Y, PARK H, CHOI B R, et al.Alteration of microbiome profile by D-allulose in amelioration of high-fat-diet-induced obesity in mice[J].Nutrients, 2020, 12(2):352.
[5] ZHANG W L, YU S H, ZHANG T, et al.Recent advances in D-allulose:Physiological functionalities, applications, and biological production[J].Trends in Food Science & Technology, 2016, 54:127-137.
[6] LEE D, HAN Y, KWON E Y, et al.D-allulose ameliorates metabolic dysfunction in C57BL/KsJ-db/db mice[J].Molecules, 2020, 25(16):3 656.
[7] MOON S, KIM Y H, CHOI K.Inhibition of 3T3-L1 adipocyte differentiation by D-allulose[J].Biotechnology and Bioprocess Engineering, 2020, 25(1):22-28.
[8] PONGKAN W, JINAWONG K, PRATCHAYASAKUL W, et al.D-allulose provides cardioprotective effect by attenuating cardiac mitochondrial dysfunction in obesity-induced insulin-resistant rats[J].European Journal of Nutrition, 2021,60(4):2 047-2 061.
[9] PRATCHAYASAKUL W, JINAWONG K, PONGKAN W, et al.Not only metformin, but also D-allulose, alleviates metabolic disturbance and cognitive decline in prediabetic rats[J].Nutritional Neuroscience, 2020.DOI:10.1080/1028415X.2020.1840050.
[10] IWASAKI Y, SENDO M, DEZAKI K, et al.GLP-1 release and vagal afferent activation mediate the beneficial metabolic and chronotherapeutic effects of D-allulose[J].Nature Communications, 2018, 9(1):113.
[11] CHUNG M Y, OH D K, LEE K W.Hypoglycemic health benefits of D-psicose[J].Journal of Agricultural & Food Chemistry, 2012, 60(4):863-869.
[12] CHATTOPADHYAY S, RAYCHAUDHURI U, CHAKRABORTY R.Artificial sweeteners—a review[J].Journal of Food Science and Technology, 2014, 51(4):611-621.
[13] ATES E G, OZVURAL E B, OZTOP M H.Understanding the role of d-allulose and soy protein addition in pectin gels[J].Journal of Applied Polymer Science, 2021, 138(8):49885.
[14] LEE P, OH H, KIM S Y, et al.Effects of d-allulose as a sucrose substitute on the physicochemical, textural, and sensorial properties of pound cakes[J].Journal of Food Processing and Preservation, 2020, 44(6):e14472.
[15] KANASAKI A, IIDA T, MURAO K, et al.D-allulose enhances uptake of HDL-cholesterol into rat′s primary hepatocyte via SR-B1[J].Cytotechnology, 2020, 72(2):295-301.
[16] TANAKA M, HAYASHI N, IIDA T, et al.Effects of chocolate containing D-allulose on postprandial lipid and carbohydrate metabolism in young Japanese women[J].Food Science and Technology Research, 2020, 26(5):623-632.
[17] MEN Y, ZHU P, ZHU Y M, et al.The development of low-calorie sugar and functional jujube food using biological transformation and fermentation coupling technology[J].Food Science & Nutrition, 2019, 7(4):1 302-1 310.
[18] 佟毅, 沈雪梅, 王靖, 等.一种新型D-阿洛酮糖3-差向异构酶及其应用:中国, CN109306347B[P].2019-08-30.
TONG Y, SHEN X M, WANG J, et al.A novel D-allulose 3-epimerase and its application:China, CN109306347B[P].2019-08-30.
[19] WEN L Q, HUANG K, WEI M H, et al.Facile enzymatic synthesis of ketoses[J].Angewandte Chemie International Edition, 2015, 54(43):12 654-12 658.
[20] BORK P, SANDER C, VALENCIA A.An ATPase domain common to prokaryotic cell cycle proteins, sugar kinases, actin, and hsp70 heat shock proteins[J].Proceedings of the National Academy of Sciences of the United States of America, 1992, 89(16):7 290-7 294.
[21] VAN DEN ENT F, LÖWE J.Crystal structure of the cell division protein FtsA from Thermotoga maritima[J].The EMBO Journal, 2000, 19(20):5 300-5 307.
[22] BUSS K A, COOPER D R, INGRAM-SMITH C, et al.Urkinase:Structure of acetate kinase, a member of the ASKHA superfamily of phosphotransferases[J].Journal of Bacteriology, 2001, 183(2):680-686.
[23] LOCHER K P, HANS M, YEH A P, et al.Crystal structure of the Acidaminococcus fermentans 2-hydroxyglutaryl-CoA dehydratase component A[J].Journal of Molecular Biology, 2001, 307(1):297-308.
[24] VAN DEN ENT F, MØLLER J J, AMOS L A, et al.F-actin-like filaments formed by plasmid segregation protein ParM[J].The EMBO Journal, 2002, 21(24):6 935-6 943.
[25] DIAO J S, COOPER D R, SANDERS D A, et al.Crystallization of butyrate kinase 2 from Thermotoga maritima mediated by vapor diffusion of acetic acid[J].Acta Crystallographica, Section D, Biological Crytallography, 2003, 59(6):1 100-1 102.
[26] MUKAI T, KAWAI S, MORI S, et al.Crystal structure of bacterial inorganic polyphosphate/ATP-glucomannokinase—insights into kinase evolution[J].Journal of Biological Chemistry, 2004, 279(48):50 591-50 600.
[27] SIMANSHU D K, SAVITHRI H S, MURTHY M R N.Crystal structures of ADP and AMPPNP-bound propionate kinase (TdcD) from Salmonella typhimurium:Comparison with members of acetate and sugar kinase/heat shock cognate 70/actin superfamily[J].Journal of Molecular Biology, 2005, 352(4):876-892.
[28] XIAO Q, NIU J R, LIU H, et al.High conversion of D-fructose into D-allulose by enzymes coupling with an ATP regeneration system[J].Molecular Biotechnology, 2019,61(6):432-441.
[29] 温俊婷. L-鼠李树胶糖激酶在D-阿洛酮糖合成中的应用[D].无锡:江南大学, 2019.
WEN J T.Application of L-rhamnulose kinase on production of D-psicose [D].Wuxi:Jiangnan University, 2019.
[30] 朱理平, 高晓冬, 李子杰, 等.一种利用全细胞转化合成D-阿洛酮糖的方法:中国, CN111518853A[P].2020-08-11.
ZHU L P, GAO X D, LI Z J, et al.A method for synthesizing D-allulose by whole cell transformation:China, CN111518853A[P].2020-08-11.
[31] MORRIS G M, HUEY R, LINDSTROM W, et al.AutoDock4 and AutoDockTools4:Automated docking with selective receptor flexibility[J].Journal of Computational Chemistry, 2009, 30(16):2 785-2 791.
[32] GRUENINGER D, SCHULZ G E.Substrate spectrum of L-rhamnulose kinase related to models derived from two ternary complex structures[J].FEBS Letters, 2007, 581(16):3 127-3 130.
[33] XIE X K, PASHKOV I, GAO X, et al.Rational improvement of simvastatin synthase solubility in Escherichia coli leads to higher whole-cell biocatalytic activity[J].Biotechnology & Bioengineering, 2009, 102(1):20-28.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.