Ribitol dehydrogenase (RDH) derived from Klebsiella oxytoca, and coding gene and application thereof

Abstract

The invention discloses a ribitol dehydrogenase (RDH) derived from Klebsiella oxytoca G4A4CGMCC No.7662, and realizes expression thereof in Escherichia coli. The experiment proves that the enzyme can realize the biotransformation production of allitol, and D-allulose can be converted into the functional rare sugar alcohol (allitol) with high conversion rate (96% or above at most) and less pollution; and an NADH (reduced nicotinamide adenine dinucleotide) regeneration system is added in the conversion process, and the NAD (nicotinamide adenine dinucleotide) consumption is low, thereby greatly lowering the cost. The allitol can be used in multiple fields such as preparation of pharmaceuticals (such as diabetes treatment medicaments and the like), health products and other raw materials of rare sugar and the like, and has wide application prospects.

Description

technical field [0001] The invention relates to an enzyme, its coding gene and its application, in particular to a ribitol dehydrogenase (RDH) derived from Klebsiella oxytoca and its application in producing allicitol. Background technique [0002] Rare sugar (Rare Sugar) is a kind of monosaccharide and its derivatives that exist in nature but are very low in content (defined by the International Rare Sugar Society ISRS in 2002). Its taste is similar to sucrose, but it has low calories, high stability, sweet It has the advantages of harmonious taste, no hygroscopicity, no cariogenicity, and high tolerance, which plays an important role in improving the diet of special populations. In addition, a large number of research results also show that rare sugars play an important physiological role in many aspects such as anti-cancer, scavenging free radicals, neuroprotection, etc., and can also undergo Maillard reactions with active substances such as proteins and peptides to optim...

AI Technical Summary

Problems solved by technology

Studies have found that some microorganisms can realize the interconversion between D-psicose and allitol, such as Enterobacter agglomerans221e (Muniruzzaman S, Tokunaga H, Izumori K. Conversion of D-psicose to allitol by Enterobacter agglomerans strain221e.J Ferment BioengVolume.1995,79(4):323-327), which reported that D-psicose was used as a substrate to transform and produce allitol, but the substrate specificity of the bacteria was poor, such as The conversion of D-fructose and D-psicose mixture may generate other by-products; another example is Enterobacter aerogenes IK7 (Gullapalli P, Takata G, Poonperm W, Rao D, Morimoto K, Akimitsu K, Tajima S, Izumori K.Bioproduction of D-psicose from allitol with Enterobacter aerogenes IK7: a new frontier in rare ketose production.Biosci Biotechnol Biochem.2007,71(12):3048-3054), the literature only reported that allitol was based on allitol The production of D-psicose by chemical transformation has not been reported in China

The main foreign patent documents are: [1] Ikumori Takeshi, Tsuzaki keiji. Ribitol dehydrogenase and its production and use. Japan, JP19940218155, 1994-08-20. This patent document discloses the production of 1- Processes for deoxygenating rare sugars such as deoxy-L-psicose and 1-deoxy-L-fructose, but not involving the production of allicitol

[2] Izumori Ken, Morimoto Kenji, Takata Goro, Tokuda Masaaki, Tsujisaka Yoshio, Takeshita Kei, Tsusaki Keiji, Okuma Kazuhiro. Microorganism with ability to produce deoxy polyol dehydrogenase and use thereof. Japan, JP20060313672, 2006-11-2 The patent literature discloses the method of using the oxidation and reduction of Enterobacter agglomerans221e to produce a variety of rare sugars and sugar alcohols respectively, but the substrate specificity of the bacteria is poor, such as the mixture of D-fructose and D-psicose Transformation may generate other by-products

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