An assay for selective determination of exo-1,4,-β-glucanases in a mixture of cellulolytic enzymes

Deshpande, Mukund V.; Eriksson, Karl‐Erik; Pettersson, L

doi:10.1016/0003-2697(84)90843-1

Cited by 290 publications

(138 citation statements)

References 16 publications

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“…P-Glucosidase activity was assayed by the method of Deshpande et al (1984) with the following modifications. The assay mixture contained 0.25 ml of diluted cell extract and 0-25 ml of 3-4 mMp-nitrophenyl P-D-glucoside in 0.05 M-phosphate/citrate buffer (pH 5.0).…”

Section: Stratagenementioning

confidence: 99%

Cloning, sequencing and analysis of expression of a Butyrivibrio fibrisolvens gene encoding a -glucosidase

Lin

Rumbak

Zappe

et al. 1990

Journal of General Microbiology

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The cloning, expression and nucleotide sequence of a 3-74 kb DNA segment on pLS215 containing a P-glucosidase gene (bglA) from Butpivibrio Jibrisolvens H17c was investigated. The B. fibrisolvens bglA open reading frame (ORF) of 2490 bp encoded a P-glucosidase of 830 amino acid residues with a calculated M, of 91800. In Escherichiu coli C6OO(pLS215) cells the P-glucosidase was localized in the cytoplasm and these cells produced an additional protein with an apparent Mr of approximately 94000. The bglA gene was expressed from its own regulatory region in E. coli and a single mRNA initiation point was identified upstream of the bglA ORF and adjacent to a promoter consensus sequence. The primary structure of the P-glucosidase showed > 40 yo similarity with a domain of 237 amino acids present in the P-glucosidases of Kluyveromyces fragilis and Clostridium thermocellum. The B. fibrisolvens P-glucosidase. hydrolysed cellobiose to a limited extent, cellotriose to cellobiose and glucose, and cellotetraose and cellopentaose to predominantly glucose.

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Section: Stratagenementioning

confidence: 99%

Cloning, sequencing and analysis of expression of a Butyrivibrio fibrisolvens gene encoding a -glucosidase

Lin

Rumbak

Zappe

et al. 1990

Journal of General Microbiology

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“…pNP -D-lactoside is known as a selective substrate digested by some cellulases. 8,[18][19][20] All these activities were induced by a cellulosic substance, and their activities in glucose medium were undetectable (data not shown). As shown in Fig.…”

Section: Resultsmentioning

confidence: 94%

Structure and Characteristics of an Endo-β-1,4-glucanase, Isolated fromTrametes hirsuta, with High Degradation to Crystalline Cellulose

Nozaki

Seki

Matsui

et al. 2007

Bioscience, Biotechnology, and Biochemistry

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Trametes hirsuta produced cellulose-degrading enzymes when it was grown in a cellulosic medium such as Avicel or wheat bran. An endo--1,4-glucanase (ThEG) was purified from the culture filtrate, and the gene and the cDNA were isolated. The gene consisted of an open reading frame encoding 384 amino acids, interrupted by 11 introns. The whole sequence showed high homology with that of family 5 glycoside hydrolase. The properties of the recombinant enzyme (rEG) in Aspergillus oryzae were compared with those of the En-1 from Irpex lacteus, which showed the highest homology among all the endoglucanases reported. The rEG activity against Avicel was about 8 times higher than that of En-1 when based on CMC degradation. A remarkable structural difference between the two enzymes was the length of the linker connecting the cellulose-binding domain to the catalytic domain.

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“…The p-nitrophenyl-β-D-cellobioside (pNPC) (Sigma, USA) was used as the substrate to assay the Tr-Cel7A activity [20]. The specific activity and the extracellular activity of Tr-Cel7A were determined by incubating the purified Tr-Cel7A protein and the supernatants collected from cell culture via centrifugation (15 min at 10,000 × g), respectively, following the described method [11].…”

Section: Assay For Tr-cel7a Activitymentioning

confidence: 99%

Promotion of Extracellular Activity of Cellobiohydrolase I from Trichoderma reesei by Protein Glycosylation Engineering in Saccharomyces cerevisiae

Shen²,

Hou³

et al. 2013

Curr Synthetic Sys Biol

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The N-glycosylation in Saccharomyces cerevisiae is of the high-mannose type, which affects the activity of the secreted heterologous glycoproteins. Cellobiohydrolase I (Tr-Cel7A) from Trichoderma reesei, is thus hyperglycosylated when expressed in S. cerevisiae. In the present work, three genes encoding the endogenous mannosyltransferases, Och1p, Mnn9p and Mnn1p, involved in glycoprotein processing in the S. cerevisiae Golgi apparatus, were individually or combinatorially disrupted to investigate the effect of the glycosylation extent on the activity of the secreted Tr-Cel7A. The glycosylation of the recombinant Tr-Cel7A was decreased and its extracellular activity was increased in all the deletion mutants. The simultaneous deletion of och1 and mnn1 has the most improvement on extracellular Tr-Cel7A activity. After expressed the α-1,2-mannosidase (Tr-Mds1p) from T. reesei in mnn1Δ/och1Δ strain, the Tr-Cel7A activity was further increased up to 320 ± 8% higher than that of the wild type strain. Such activity improvement was due not only to the higher secretion yield but also to the increased specific activity resulted from the changes in glycosylation. The results thus indicated that protein glycosylation engineering in S. cerevisiae was an effective approach to improve the extracellular activity of Tr-Cel7A.

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An assay for selective determination of exo-1,4,-β-glucanases in a mixture of cellulolytic enzymes

Cited by 290 publications

References 16 publications

Cloning, sequencing and analysis of expression of a Butyrivibrio fibrisolvens gene encoding a -glucosidase

Cloning, sequencing and analysis of expression of a Butyrivibrio fibrisolvens gene encoding a -glucosidase

Structure and Characteristics of an Endo-β-1,4-glucanase, Isolated fromTrametes hirsuta, with High Degradation to Crystalline Cellulose

Promotion of Extracellular Activity of Cellobiohydrolase I from Trichoderma reesei by Protein Glycosylation Engineering in Saccharomyces cerevisiae

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