Methods for measuring cellulase activities

Wood, Thomas M.; Bha, K M

doi:10.1016/0076-6879(88)60109-1

Cited by 1,175 publications

(642 citation statements)

References 21 publications

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“…nlm .ni h .gov) , Enzymic assays and detection of hydrolysis products. Enzyme aliquots in standard assays were incubated in sodium phosphate/citrate buffer (50 mM, pH 6.2) with 0.7 '/o substrate at 85 "C. Reducing sugars released from polymeric substrates were detected by the 3,s-dinitrosalicylic acid method (Wood & Bhat, 1988). One unit of activity is defined by the release of 1 pmol glucose equivalent min-l. Digests of laminarin were applied to TLC (Silica-gel plates).…”

Section: Methodsmentioning

confidence: 99%

Highly thermostable endo-1,3-β -glucanase (laminarinase) Lam A from Thermotoga neapolitana: nucleotide sequence of the gene and characterization of the recombinant gene product

et al. 1997

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The nucleotide sequence of clone p n 2 6 (3786 bp), containing the gene for 1,3-pglucanase LamA (laminarinase) from Tnermofoga neapolitana, was determined. It contains an ORF encoding a protein of 646 aa (73328 Da). The central part of the protein is homologous to the complete catalytic domain of bacterial and some eukaryotic endo-l,3-&~-glucanases and belongs to family 16 of glycosyl hydrolases. This domain is flanked on both sides by one copy on each side of a substrate binding domain homologue (family 11). The recombinant laminarinase protein was purified from Escherichia coli host cells in two forms, a 73 kDa and a processed 52 kDa protein, both having high specific activity towards laminarin (3100 and 2600 U mg-l, respectively) and Km values of 2.8 and 2.2 mg mF, respectively. Limited activity on 1,3-1,4-/3-glucan (lichenan) was detected (90 U mg-l). Laminarin was degraded in an endoglucanase modus, yielding glucose, laminaribiose and ltriose as end products. Thus LamA classifies as an endo-l,3(4)-/3=gIucanase (EC 3.2.1.6). The optimum temperature of the enzymes was 95 "C (73 kDa) and 85 "C (52 kDa) at an optimum pH of 62. The superior thermostability of the 73 kDa enzyme is demonstrated by incubation without substrate at 100 "C, where 57% of the initial activity remained after 30 min (82% at 95 OC). Thus, LamA is the most thermostable 1,3-&glucanase described to date.

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Section: Methodsmentioning

confidence: 99%

Highly thermostable endo-1,3-β -glucanase (laminarinase) Lam A from Thermotoga neapolitana: nucleotide sequence of the gene and characterization of the recombinant gene product

et al. 1997

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“…The total cellulolitic activity (FPase) was determined by the standard method (Ghose, 1987) and the glucose released was determined by the DNS method described by Miller (1959). The activity of the enzyme β-glucosidade was determined by the methodology described by Wood and Bhat (1988).…”

Section: Enzymatic Hydrolysismentioning

confidence: 99%

Assessment of sugarcane trash for agronomic and energy purposes in Brazil

Franco

Pimenta

Carvalho

et al. 2013

Sci. agric. (Piracicaba, Braz.)

108

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Due to new possibilities for using sugarcane (Saccharum spp.) trash for electricity generation, and the production of 2 nd generation ethanol and others chemicals, the interest for its recovery has increased. However, the question of how much trash can be removed from sugarcane field still needs to be clarified. This study evaluated the amount of dry matter, nutrients content, structural compounds and efficiency of the enzymatic hydrolysis of the hydrothermal pretreated materials for tops and dry leaves in samples from sugarcane varieties. Tops and dry leaves present differences in nutrients content and moisture. Therefore, the amount of trash to be collected should not be simply based on percentages, but also should take into account the different fractions of the crop residues. For instance, around 80 % of N, P and K were derived from tops. Therein, the environmental indicators of the entire chain of sugarcane could be benefited because more nutrients would be recycled and less mineral fertilizers might be used for sugarcane production if tops are left on the field. Further, the tops have seven times more moisture than dry leaves and higher amounts of extractives (organic compounds of low molecular weight). Moreover, as the result of yield obtained in the pretreatment steps for dry leaves were superior to the tops and the glucose yields obtained in the enzymatic hydrolysis step were similar, it can be predicted that for second generation ethanol production, it is more viable to recover parts of the dry leaves fraction, leaving the tops on the field.

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“…Lignin peroxidase activity was monitored at pH 3.0 according to Tien & Kirk (1988), and the formation of veratraldehyde was monitored at 310 nm (ε310 = 9.3 mM -1 cm -1 ). Laccase activity was determined according to Dias et al (2003) by measuring the oxidation of 2mM 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonate (ABTS) buffered with 100 mM citrate-phosphate (pH 4.0) and formation of For cellulolytic enzyme assays, activities of carboxymethylcellulase (CMCase) and Avicell digesting cellulase (avicelase) were measured according to the IUPAC (Wood and Bhat, 1988). The reducing sugars released were determined by the dinitrosalicylic acid (DNS), using glucose as a standard (Bezerra and Dias, 2004).…”

Section: Enzyme Assaysmentioning

confidence: 99%