An extremely highly active cellobiohydrolase (CBH IIb or Cel6B) was isolated from Chrysosporium lucknowense UV18-25 culture filtrate. The CBH IIb demonstrated the highest ability for a deep degradation of crystalline cellulose amongst a few cellobiohydrolases tested, including C. lucknowense CBH Ia, Ib, IIa, and Trichoderma reesei CBH I and II. Using purified C. lucknowense enzymes (CBH Ia, Ib, and IIb; endoglucanases II and V; beta-glucosidase, xylanase II), artificial multienzyme mixtures were reconstituted, displaying an extremely high performance in a conversion of different cellulosic substrates (Avicel, cotton, pretreated Douglas fir wood) to glucose. These mixtures were much or notably more effective in hydrolysis of the cellulosic substrates than the crude multienzyme C. lucknowense preparation and other crude cellulase samples produced by T. reesei and Penicillium verruculosum. Highly active cellulases are a key factor in bioconversion of plant lignocellulosic biomass to ethanol as an alternative to fossil fuels.

Using different chromatographic techniques, eight cellulolytic enzymes were isolated from the culture broth of a mutant strain of Chrysosporium lucknowense: six endoglucanases (EG: 25 kD, pI 4.0; 28 kD, pI 5.7; 44 kD, pI 6.0; 47 kD, pI 5.7; 51 kD, pI 4.8; 60 kD, pI 3.7) and two cellobiohydrolases (CBH I, 65 kD, pI 4.5; CBH II, 42 kD, pI 4.2). Some of the isolated cellulases were classified into known families of glycoside hydrolases: Cel6A (CBH II), Cel7A (CBH I), Cel12A (EG28), Cel45A (EG25). It was shown that EG44 and EG51 are two different forms of one enzyme. EG44 seems to be a catalytic module of an intact EG51 without a cellulose-binding module. All the enzymes had pH optimum of activity in the acidic range (at pH 4.5-6.0), whereas EG25 and EG47 retained 55-60% of the maximum activity at pH 8.5. Substrate specificity of the purified cellulases against carboxymethylcellulose (CMC), beta-glucan, Avicel, xylan, xyloglucan, laminarin, and p-nitrophenyl-beta-D-cellobioside was studied. EG44 and EG51 were characterized by the highest CMCase activity (59 and 52 U/mg protein). EG28 had the lowest CMCase activity (11 U/mg) amongst the endoglucanases; however, this enzyme displayed the highest activity against beta-glucan (125 U/mg). Only EG51 and CBH I were characterized by high adsorption ability on Avicel cellulose (98-99%). Kinetics of Avicel hydrolysis by the isolated cellulases in the presence of purified beta-glucosidase from Aspergillus japonicus was studied. The hydrolytic efficiency of cellulases (estimated as glucose yield after a 7-day reaction) decreased in the following order: CBH I, EG60, CBH II, EG51, EG47, EG25, EG28, EG44.

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SummaryBased on surveys during 2000–2012 and best available knowledge, we estimate the global population size of the Endangered Scaly-sided Merganser Mergus squamatus to be c.1,940 pairs (or c.4,660 birds prior to reproduction). In Russia, surveys identified the breeding range in the Sikhote-Alin mountain range and the area adjoining the lower Amur River. Particular effort was made to define the edge of the range. Breeding densities in the Sikhote-Alin were applied to un-surveyed rivers within the range to estimate overall numbers. The breeding range comprises c.7,800 km of 120 rivers: 6,800 km in Russia, 600 in China and 400 in DPR Korea. In Russia it comprises 88 rivers of both the western and eastern slopes of the Sikhote-Alin Range, and two small isolated areas, one in central Khabarovsk and the other in the Jewish Autonomous Oblast. All known and potential breeding areas were surveyed in China, where the breeding range comprises the western slope of the Changbai Mountains, and one isolated river system in the Lesser Xingan Mountains. Scaly-sided Mergansers are also assumed to breed on the eastern slope of the Changbai Mountains in DPR Korea. No recent surveys have been made there but numbers were estimated by applying densities from China to rivers with apparently suitable habitat. Detailed examination of past records suggests that an earlier claim of breeding Scaly-sided Mergansers on the Dep River was erroneous and we conclude that the breeding range never extended as far west as has been portrayed in most literature. The lack of comparable surveys previously precludes an assessment of change in population range or size, though it seems inevitable that habitat loss in China is likely to have resulted in some loss of range and numbers, particularly in Heilongjiang Province, in recent decades.

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