عنوان

Extracellular beta-D -mannanase activity from Trichoderma harzianum E58

شماره

TLpq89225670

زبان متن نوشتاري يا گفتاري و مانند آن

انگلیسی

عنوان اصلي

Extracellular beta-D -mannanase activity from Trichoderma harzianum E58

نام عام مواد

[Thesis]

نام نخستين پديدآور

J. P. Torrie

نام ساير پديدآوران

J. N. Saddler

نام ناشر، پخش کننده و غيره

University of Ottawa (Canada)

تاریخ نشرو بخش و غیره

1991

نام خاص و کميت اثر

242

جزئيات پايان نامه و نوع درجه آن

Ph.D.

کسي که مدرک را اعطا کرده

University of Ottawa (Canada)

امتياز متن

1991

متن يادداشت

In this work, 4 yeasts (Pichia wickerhammi, Candida wickerhammi, Pichia stipitis CBS5776, Pichia stipitis CBS5876), and 5 fungi (Theivalia spp., Thermoascus aurantiacus, Tyromyces palustris A, Aspergillus niger, Trichoderma harzianum), known to excrete enzymes capable of hydrolyzing polysaccharides found in association with usd\betausd-D-mannans, were assessed for their ability to degrade usd\betausd-D-mannans. The usd\betausd-D-mannanase activity found in the culture filtrate of T. harzianum was selected for further study and a 'cellulase-free' usd\betausd-D-mannanase isolated from culture filtrates of T. harzianum grown on medium supplemented with 1% w/v locust bean gum studied. usd\betausd-D-Mannanase activity was detected in T. harzianum culture filtrates from media supplemented with 1% (w/v) Avicel, locust bean gum galactomannan, konjac root glucomannan, or spruce wood water-solubles. Medium supplemented with 1% (w/v) mannose did not induce usd\betausd-D-glucomannanase or usd\betausd-D-galactomannanase activity. However, when 0.5% (w/v) usd\betausd-D-galactomannan was added with mannose, usd\betausd-D-mannanase activity was detected in the culture filtrate. Growth of the fungus on mannan-rich locust bean gum resulted in the highest specific usd\betausd-D-glucomannanase and usd\betausd-D-galactomannanase values. A zymogram assay was developed to selectively detect usd\betausd-D-mannanase activity in crude culture filtrates. The presence of different polysaccharides in the growth medium resulted in different usd\betausd-D-mannanase zymogram profiles. Analyses of the protein profiles of the culture filtrates separated by isoelectric focusing revealed several bands having usd\betausd-D-mannanase and endoglucanase activity. A protein band having usd\betausd-D-mannanase activity but lacking detectable cellulase activity was identified. This enzyme was purified to homogeneity via a sequence involving ultrafiltration, ion exchange and gel filtration. This 'cellulase-free' usd\betausd-D-mannanase has the highest reported pI for a fungal usd\betausd-D-mannanase. The isolated enzyme had a molecular weight of 42.9 4 kD, an optimum temperature of 60-65C, an optimum pH of 5.8, a pI of 6.55, and possessed at least 75% of maximum activity over a pH range from 3.21-6.8. Enzymatic activity was stable during 12 months of storage at 4C. usd\betausd-D-mannanase activity was resistant to pepsin, usd\alphausd-chymotrypsin, trypsin, and Staphylococcus V8 protease. The effect of metal ions, detergent and solvents on usd\betausd-D-mannanase activity was also determined. Although the enzyme did not degrade Avicel or Solka floc, it did associate with both celluloses. Enzyme associated with these celluloses remained active towards locust bean gum galactomannan. The overall efficiency of the enzyme (Vmax/K{\rm m}) for the target substrate, locust bean gum galactomannan, was reduced by the presence of 1.0% w/v Avicel. Of the four usd\betausd-D-mannans tested, the enzyme had greatest overall efficiency towards konjac glucomannan. However, deacetylation of konjac glucomannan lowered the efficiency of the enzyme by 41.5%. (Abstract shortened by UMI.)

موضوع مستند نشده

Biochemistry

موضوع مستند نشده

mannanase

موضوع مستند نشده

Pure sciences

مستند نام اشخاص تاييد نشده

J. N. Saddler

مستند نام اشخاص تاييد نشده

J. P. Torrie

نام الکترونيکي

فرمت انتشار

p

نوع ماده

[Thesis]

کد کاربرگه

276903

سطح دسترسي

a

تكميل شده

Y