Purification and characterization of a novel thermostable <i>β</i>-amylase from <i>Clostridium thermosulphurogenes</i>

Shen, Gwo‐Jenn; Saha, Badal C.; Lee, Yong‐Eok; Bhatnagar, L.; Zeikus, J. G.

doi:10.1042/bj2540835

Cited by 64 publications

(40 citation statements)

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“…Advantages of using these thermostable and thermophilic enzymes in industrial processes were proposed (Zeikus, 1979;Ng & Kenealy, 1987). Our laboratory has reported on purification and biochemical characterization of thermostable fl-amylase and amylopullulanase from Clostridium thermosulfurogenes and C. thermohydrosulfuricum respectively Shen et al, 1988). In spite of the large number of studies on glucose isomerases from various enzyme sources, nothing is known about the detailed biochemical or molecular-genetic properties of glucose isomerases from thermoanaerobic bacteria.…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of thermostable glucose isomerase from Clostridium thermosulfurogenes and Thermoanaerobacter strain B6A

Lee

Zeikus

1991

Biochemical Journal

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Glucose isomerases produced by Thermoanaerobacter strain B6A and Clostridium thermosulfurogenes strain 4B were purified 10-11-fold to homogeneity and their physicochemical and catalytic properties were determined. Both purified enzymes displayed very similar properties (native Mr 200000, tetrameric subunit composition, and apparent pH optima 7.0-7.5). The enzymes were stable at pH 5.5-12.0, and maintained more than 90% activity after incubation at high temperature (85°C) for 1 h in the presence of metal ions. The N-terminal amino acid sequences of both thermostable glucose isomerases were Met-Asn-Lys-Tyr-Phe-Glu-Asn and were not similar to that of the thermolabile Bacillus subtilis enzyme. The glucose isomerase from C. thermosulfurogenes and Thermoanaerobacter displayed pI values of 4.9 and 4.8, and their k,,t and Km values for D-glucose at 65°C were 1040 and 1260 min-' and 140 and 120 mm respectively. Both enzymes displayed higher kc.t and lower Km values for D-xylose than for D-glucose. The C. thermosulfurogenes enzyme required Co2+ or Mg2+ for thermal stability and glucose isomerase activity, and Mn2+ or these metals for xylose isomerase activity. Crystals of C. thermosulfurogenes glucose isomerase were formed at room temperature by the hanging-drop method using 16-180% poly(ethylene glycol) (PEG) 4000 in 0.1 M-citrate buffer.

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Section: Introductionmentioning

confidence: 99%

Purification and characterization of thermostable glucose isomerase from Clostridium thermosulfurogenes and Thermoanaerobacter strain B6A

Lee

Zeikus

1991

Biochemical Journal

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“…Thermostable exo-acting β -amylases and glucoamylases are also available. β-amylases have for example been isolated from Thermotoga maritima [58] and Clostridium thermosulfurigenes [59,60]. The T. maritima enzyme has optimal activity at 95 °C and pH 5, while the C. thermosulfurigenes is optimally active at 75 °C, pH 5 although its stability is enhanced by Ca 2+ .…”

Section: Major Bottlenecksmentioning

confidence: 99%

Thermostable Glycoside Hydrolases in Biorefinery Technologies

Linares-Pastén¹,

Andersson²,

Karlsson

2014

CBIOT

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Glycoside hydrolases, which are responsible for the degradation of the major fraction of biomass, the polymeric carbohydrates in starch and lignocellulose, are predicted to gain increasing roles as catalysts in biorefining applications in the future bioeconomy. In this context, thermostable variants will be important, as the recalcitrance of these biomass-components to degradation often motivates thermal treatments. The traditional focus on degradation is also predicted to be changed into more versatile roles of the enzymes, also involving specific conversions to defined products. In addition, integration of genes encoding interesting target activities opens the possibilities for whole cell applications, in organisms allowing processing at elevated temperatures for production of defined metabolic products.In this review, we overview the application of glycoside hydrolases related to the biorefining context (for production of food, chemicals, and fuels). Use of thermostable enzymes in processing of biomass is highlighted, moving from the activities required to act on different types of polymers, to specific examples in today's processing. Examples given involve (i) monosaccharide production for food applications as well as use as carbon source for microbial conversions (to metabolites such as fuels and chemical intermediates), (ii) oligosaccharide production for prebiotics applications (iii) treatment for plant metabolite product release, and (iv) production of surfactants of the alkyl glycoside class. Finally future possibilities in whole cell biorefining are shown.

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“…The enzyme showed remarkable thermostability being fully active at 121°C for 15 min. Previously, a thermostable b amylase was isolated from Clostridium thermosulfurogenes by Shen et al (1988) that was stable at 80°C. Since thermostability is the crucial criteria for industrial application (Haki and Rakshit 2003), b amylase of this study has got a leading edge over all previously reported b amylase.…”

Section: Introductionmentioning

confidence: 99%

Optimization of novel hyperthermostable β amylase production by Bacillus subtilis DJ5 using solid agroresidual substrates

Poddar

Jana

2013

Int. J. Environ. Sci. Technol.

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Eight different low cost starchy agroresidues namely Barley (B), Wheat bran (WB), Sattu (S), Rice powder (RP), Corn flour (CF), Rice husk (RH), Yellow peas split (YPS) and arrowroot (A) were used for solid culturing of Bacillus subtilis DJ5 for production of novel hyperthermostable b amylase. Various process parameters like initial moisture content, inoculum load, medium pH and incubation temperature affecting enzyme production were optimized to ensure maximum enzyme yield. Only 10 % inoculums load and medium pH of 6.9 was found sufficient to achieve maximum enzyme production in all substrates in a decreasing order,Optimum b amylase production was highly dependent on initial moisture content of substrate as observed from varying requirement of moisture for different substrates. Only 50 % moisture was sufficient for maximum enzyme production of 84.29 U/gdm in CF. For B, RH, YPS, and A 60 % initial moisture resulted in higher production of 120.34, 35.19, 26.59, and 21.58 U/ gdm, respectively, at 37°C. However, for S and RP higher (70 %) moisture content allowed 113.4 and 85.56 U/gdm enzyme production, respectively. Under optimized conditions, maximum b amylase production was observed after 25 h for A, YPS, RH, RP; 41 h for B, WB, CF, and 45 h for S.

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Purification and characterization of a novel thermostable β-amylase from Clostridium thermosulphurogenes

Cited by 64 publications

References 20 publications

Purification and characterization of thermostable glucose isomerase from Clostridium thermosulfurogenes and Thermoanaerobacter strain B6A

Purification and characterization of thermostable glucose isomerase from Clostridium thermosulfurogenes and Thermoanaerobacter strain B6A

Thermostable Glycoside Hydrolases in Biorefinery Technologies

Optimization of novel hyperthermostable β amylase production by Bacillus subtilis DJ5 using solid agroresidual substrates

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