Isolation and characterization of oligosaccharides produced from sucrose by transglucosylation action of Serratia plymuthica.

Fujii, Satoshi; Kishihara, Shiro; Kômoto, Masahiko; Shimizu, Jun-ichi

doi:10.3136/nskkk1962.30.339

Cited by 21 publications

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“…Several microorganisms, such as Protaminobacter rubrum (32), Serratia plymuthica (11), Klebsiella planticola (18), and Erwinia rhapontici (5), have been found to form palatinose and trehalulose from sucrose. The adaptive role of sucrose isomer formation is unclear.…”

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confidence: 99%

Cloning and Characterization of the Gene Cluster for Palatinose Metabolism from the Phytopathogenic BacteriumErwinia rhapontici

2001

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Erwinia rhapontici is able to convert sucrose into isomaltulose (palatinose, 6-O-␣-D-glucopyranosyl-D-fructose) and trehalulose (1-O-␣-D-glucopyranosyl-D-fructose)by the activity of a sucrose isomerase. These sucrose isomers cannot be metabolized by plant cells and most other organisms and therefore are possibly advantageous for the pathogen. This view is supported by the observation that in vitro yeast invertase activity can be inhibited by palatinose, thus preventing sucrose consumption. Due to the lack of genetic information, the role of sucrose isomers in pathogenicity has not been evaluated. Here we describe for the first time the cloning and characterization of the palatinose (pal) genes from Erwinia rhapontici. To this end, a 15-kb chromosomal DNA fragment containing nine complete open reading frames (ORFs) was cloned. The pal gene products of Erwinia rhapontici were shown to be homologous to proteins involved in uptake and metabolism of various sugars from other microorganisms. The palE, palF, palG, palH, palK, palQ, and palZ genes were oriented divergently with respect to the palR and palI genes, and sequence analysis suggested that the first set of genes constitutes an operon. Northern blot analysis of RNA extracted from bacteria grown under various conditions implies that the expression of the palI gene and the palEFGHKQZ genes is oppositely regulated at the transcriptional level. Genes involved in palatinose uptake and metabolism are down regulated by sucrose and activated by palatinose. Palatinose activation is inhibited by sucrose. Functional expression of palI and palQ in Escherichia coli revealed sucrose isomerase and palatinase activity, respectively.Isomaltulose (commonly referred to as palatinose, 6-O-␣-Dglucopyranosyl-D-fructose) and trehalulose (1-O-␣-D-glucopyranosyl-D-fructose) are functional isomers of sucrose. Several microorganisms, such as Protaminobacter rubrum (32), Serratia plymuthica (11), Klebsiella planticola (18), and Erwinia rhapontici (5), have been found to form palatinose and trehalulose from sucrose. The adaptive role of sucrose isomer formation is unclear. Microorganisms in nature are often faced with a "feast-or-famine" type of existence, and many bacteria have evolved biochemical systems for the production of storage compounds that serve as reserve material. These storage compounds become especially important under conditions of limited nutrient supply. Therefore, it was suspected that the bioconversion of sucrose may be a method of irreversibly sequestering a carbon and energy source in a form unavailable to competitors such as the host plant or other microorganisms (6), and sucrose conversion by Erwinia rhapontici may play a similar role to 3-ketosucrose formed by Agrobacterium tumefaciens (12) and the gluconate and 2-oxoglucose produced by Pseudomonas aeruginosa (33).The formation of sucrose isomers in E. rhapontici, a pathogen associated with crown rot in rhubarb, is accomplished through the activity of a single enzyme, which has been located to the cell's periplasmi...

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Cloning and Characterization of the Gene Cluster for Palatinose Metabolism from the Phytopathogenic BacteriumErwinia rhapontici

2001

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“…Isomaltulose synthase has been studied by many investigators from the point of intramolecular transglycosylation. Protaminobacter rubrum, 9,1 0) Serratia plymutica, [10][11][12] Erwinia, 13,14) and Klebsiella 15 ) are known to produce this enzyme. Isomaltulose synthase catalyzes the conversion of sucrose into the structural isomers, isomaltulose (paratinose, 6-0-I1-o-glucopyranosyl-o-fructofuranose) and trehalulose (l-O-I1-o-glucopyranosyl-o-fructofuranose) simultaneously.…”

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confidence: 99%

Purification and Properties of a Novel Enzyme, Maltooligosyl Trehalose Synthase, fromArthrobactersp. Q36

Nakada

Maruta

Tsusaki

et al. 1995

Bioscience, Biotechnology, and Biochemistry

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“…A isomaltulose é também utilizada na produção de isomalte, uma mistura de açúcar álcool, de baixo valor calórico e baixo potencial cariogênico. No estudo da influência dos componentes do meio de cultivo, na produção de glicosiltransferase, em frascos agitados, foi obtido maior atividade da enzima (12,8 unidades de atividade/mL do meio de cultura) em meio de cultura A constituído de melaço 12% (p/v) de sólidos solúveis totais, peptona 4% (p/v) e extrato de carne 0,4% (p/v). No estudo do efeito do tempo e da temperatura na fermentação da linhagem de Erwinia sp D12, em fermentador New Brunswick de 3L, contendo meio de cultura A, foi obtida maior atividade de glicosiltransferase (15,6 unidades de atividade/ mL de meio de cultura) na fase exponencial de crescimento após 8 horas de fermentação a 30ºC.…”

Section: Introductionunclassified

Produção de isomaltulose a partir da transformação enzimática da sacarose, utilizando-se Erwinia sp D12 imobilizada com alginato de cálcio

Moraes¹,

Steckelberg²,

Sato³

et al. 2005

Ciênc. Tecnol. Aliment.

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-INTRODUÇÃOAtravés de métodos biotecnológicos e enzimáticos é possível obter monossacarídeos, dissacarídeos, oligossacarídeos e polissacarídeos derivados da sacarose, de importância para a indústria de alimentos e farmacêutica. Uma das transformações possíveis consiste na conversão da sacarose em isomaltulose através do uso de glicosiltransferase microbiana [12].A isomaltulose (6-0-α-D-glicopiranosil-D-frutofuranose) é um dissacarídeo redutor, isômero estrutural da sacarose (2-0-α-D-glicopiranosil-D-frutofuranose), encontrado naturalmente no mel e no caldo de canade-açúcar. A isomaltulose, também conhecida como palatinose, possui um sabor adocicado suave e propriedades físicas e organolépticas muito similares às da sacarose. A solubilidade da isomaltulose é aproximadamente a metade da sacarose, o que facilita a sua separação na etapa de cristalização [11].Devido ao baixo potencial cariogênico, a isomaltulose é utilizada comercialmente na produção de doces e gomas de mascar. A cárie dentária é a mais prevalente doença entre a população humana civilizada. A microbiota oral é constituída de microrganismos que geram ácidos através da degradação de carboidratos, levando a formação das cáries. A produção de ácidos pelo Streptococcus mutans é menor a partir de isomaltulose, quando comparado com a utilização da sacarose. A isomaltulose é também empregada na produção de isomalte, um açúcar-álcool formado por quantidades equimolares de dois isômeros SUMMARY PRODUCTION OF ISOMALTULOSE FROM ENZYMATIC TRANSFORMATION OF SUCROSE, USINGErwinia sp D12 IMMOBILIZED WITH CALCIUM ALGINATE. The glucosyltransferase of Erwinia sp D12 is able to convert sucrose into isomaltulose (6-0-α-D-glucopyranosyl-Dfructofuranose), an alternative sugar which presents low cariogenic potential, and can be used to produce chocolate, chewing gum and candy. The isomaltulose is also used to produce isomalt, a mixture of alcohol-sugar with a low caloric value and low cariogenicity power.In the study of the influence of the components of the culture medium in the glucosyltransferase production in flasks under shaking conditions, the highest activity (12.8 units of activity /mL of culture medium) was obtained in culture medium A, containing molasses 12% (p/v) of the total soluble solid, peptone 4% (p/v) and meat extract 0,4% (p/v). In the study of the effects of time and temperature on the fermentation of Erwinia sp D12 in a 3L New Brunswick fermentor containing culture medium A. the highest glucosyltransferase activity (15.6 units of activity /mL of culture medium) was obtained during the exponential growth phase after 8 hours of fermentation at 30ºC. In the production of isomaltulose from enzymatic transformation of sucrose by Erwinia sp D12 cells immobilized in calcium alginate, the effects of the temperature (25-35ºC) and substrate concentration (12,5-60%) were evaluated, the yield of isomaltulose was approximately 50%, from sucrose solutions ranging from 20 to 30% at 35ºC. Excess of sucrose affected the activity of the immobilized cell, decreasing con...

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Isolation and characterization of oligosaccharides produced from sucrose by transglucosylation action of Serratia plymuthica.

Cited by 21 publications

References 0 publications

Cloning and Characterization of the Gene Cluster for Palatinose Metabolism from the Phytopathogenic BacteriumErwinia rhapontici

Cloning and Characterization of the Gene Cluster for Palatinose Metabolism from the Phytopathogenic BacteriumErwinia rhapontici

Purification and Properties of a Novel Enzyme, Maltooligosyl Trehalose Synthase, fromArthrobactersp. Q36

Produção de isomaltulose a partir da transformação enzimática da sacarose, utilizando-se Erwinia sp D12 imobilizada com alginato de cálcio

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