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Jason C. H. Shih

Bio: Jason C. H. Shih is an academic researcher from North Carolina State University. The author has contributed to research in topics: Keratinase & Bacillus licheniformis. The author has an hindex of 25, co-authored 58 publications receiving 2531 citations.


Papers
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Journal ArticleDOI
TL;DR: The purified keratinase hydrolyzes a broad range of substrates and displays higher proteolytic activity than most proteases and is a useful enzyme for promoting the hydrolysis of feather keratin and improving the digestibility of feather meal.
Abstract: A keratinase was isolated from the culture medium of feather-degrading Bacillus licheniformis PWD-1 by use of an assay of the hydrolysis of azokeratin. Membrane ultrafiltration and carboxymethyl cellulose ion-exchange and Sephadex G-75 gel chromatographies were used to purify the enzyme. The specific activity of the purified keratinase relative to that in the original medium was approximately 70-fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and Sephadex G-75 chromatography indicated that the purified keratinase is monomeric and has a molecular mass of 33 kDa. The optimum pH and the pI were determined to be 7.5 and 7.25, respectively. Under standard assay conditions, the apparent temperature optimum was 50°C. The enzyme is stable when stored at −20°C. The purified keratinase hydrolyzes a broad range of substrates and displays higher proteolytic activity than most proteases. In practical applications, keratinase is a useful enzyme for promoting the hydrolysis of feather keratin and improving the digestibility of feather meal. Images

363 citations

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TL;DR: A feather-degrading culture was enriched with isolates from a poultry waste digestor and adapted to grow with feathers as its primary source of carbon, sulfur, and energy, indicating a potential biotechnique for degradation and utilization of feather keratin.
Abstract: A feather-degrading culture was enriched with isolates from a poultry waste digestor and adapted to grow with feathers as its primary source of carbon, sulfur, and energy. Subsequently, a feather-hydrolytic, endospore-forming, motile, rod-shaped bacterium was isolated from the feather-degrading culture. The organism was Gram stain variable and catalase positive and demonstrated facultative growth at thermophilic temperatures. The optimum rate of growth in nutrient broth occurred at 45 to 50°C and at pH 7.5. Electron microscopy of the isolate showed internal crystals. The microorganism was identified as Bacillus licheniformis PWD-1. Growth on hammer-milled-feather medium of various substrate concentrations was determined by plate colony count. Maximum growth (approximately 109 cells per ml) at 50°C occurred 5 days postinoculation on 1% feather substrate. Feather hydrolysis was evidenced as free amino acids produced in the medium. The most efficient conditions for feather fermentation occurred during the incubation of 1 part feathers to 2 parts B. licheniformis PWD-1 culture (107 cells per ml) for 6 days at 50°C. These data indicate a potential biotechnique for degradation and utilization of feather keratin.

335 citations

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TL;DR: Results indicate that the anaerobic fermentation of feathers offers a potential new process for feather waste treatment to provide a nutritious feed protein.

162 citations

Journal ArticleDOI
TL;DR: Results indicated that only in the presence of detergents did heat pretreatment at >100 degrees C allow the extensive enzymatic breakdown of PrPSc to a state where it is immunochemically undetectable, which could lead to the development of a method for the decontamination of medical and laboratory equipment.
Abstract: Prions-infectious agents involved in transmissible spongiform encephalopathies-normally survive proteolytic and mild protein-destructive processes. Using bacterial keratinase produced by Bacillus licheniformis strain PWD-1, we tested conditions to accomplish the full degradation of prion protein (PrP) in brain-stem tissue from animals with bovine spongiform encephalopathy and scrapie. The detection of PrPSc, the disease-associated isoform of PrP, in homogenates was done by Western blotting and various antibodies. The results indicated that only in the presence of detergents did heat pretreatment at >100 degrees C allow the extensive enzymatic breakdown of PrPSc to a state where it is immunochemically undetectable. Proteinase K and 2 other subtilisin proteases, but not trypsin and pepsin, were also effective. This enzymatic process could lead to the development of a method for the decontamination of medical and laboratory equipment. The ultimate effectiveness of this method of prion inactivation has to be tested in mouse bioassays.

153 citations

Journal ArticleDOI
TL;DR: NorthernRNA analysis demonstrates that transcriptional regulation controls kerA expression on different growth media and deduced amino acid sequences indicate only three amino acid differences between the two mature proteases.
Abstract: Bacillus licheniformis PWD-1 (ATCC 53757) secretes keratinase, a proteolytic enzyme which is active on whole feathers. By amino acid sequence similarity and phenylmethylsulfonyl fluoride inhibition, the keratinase was demonstrated to be a serine protease. The entire nucleotide sequence of the coding and flanking regions of the keratinase structure gene, kerA, was determined. A fixed oligonucleotide primer derived from the N-terminal sequence of the purified enzyme and a second random oligonucleotide primer were used in a procedure called PCR walking, which was developed to amplify and sequence the upstream and downstream regions of kerA. Another method, PCR screening, was conducted with a lambda phage vector with inserted PWD-1 genomic DNA fragments as templates and with the known sequences of the vector arms and the N-terminal sequence of the enzyme as primers. PCR amplification and sequence analysis of the lambda library completed the entire kerA sequence and established a set of gene deletions. The kerA gene shares a 97% sequence identity with the gene encoding subtilisin Carlsberg from B. licheniformis NCIMB 6816. The putative promoters, ribosome binding sites, and transcriptional terminators are also similar in these two bacteria. The deduced amino acid sequences indicate only three amino acid differences between the two mature proteases. Northern (RNA) analysis demonstrates that transcriptional regulation controls kerA expression on different growth media.

147 citations


Cited by
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Journal ArticleDOI
01 Mar 1941-Nature
TL;DR: In this article, Gray has written a book on diseases of poultry, Diseases of Poultry Their Aetiology, Diagnosis, Treatment and Control; with a Section on the Normal Anatomy and Physiology of the Fowl.
Abstract: VERY few veterinary surgeons have thought fit to write a book on diseases of poultry. Mr. Ernest Gray has done justice to the subject and is to be congratulated on his effort. A book of this size, written by one with specialized knowledge, will add to the value of any library or private bookshelf. Diseases of Poultry Their Aetiology, Diagnosis, Treatment and Control; with a Section on the Normal Anatomy and Physiology of the Fowl. By Ernest Gray. (Lockwood's Agricultural and Horticultural Handbooks.) Pp. x + 198 + 16 plates. (London: Crosby Lockwood and Son, Ltd., 1940.) 9s. 6d. net.

1,282 citations

Journal ArticleDOI
TL;DR: Developments from the understanding of the complex protein translocation machinery of Gram-positive bacteria should allow the resolution of current secretion challenges and make Bacillus species preeminent hosts for heterologous protein production.
Abstract: Bacillus species continue to be dominant bacterial workhorses in microbial fermentations. Bacillus subtilis (natto) is the key microbial participant in the ongoing production of the soya-based trad...

1,100 citations

Journal ArticleDOI
TL;DR: In this paper, a review of the proteases that can resist extreme alkaline environments produced by a wide range of alkalophilic microorganisms is presented, and various nutritional and environmental parameters affecting the production of alkaline proteases are delineated.

806 citations

Journal ArticleDOI
TL;DR: Information pertaining to the utilization of by-products and waste materials from meat, poultry and fish and their processing industries has been reviewed here and Medicinal and pharmaceutical uses ofBy-product are highlighted in this review.
Abstract: India is bestowed with vast livestock wealth and it is growing at the rate of 6% per annum. The contribution of livestock industry including poultry and fish is increasing substantially in GDP of country which accounts for >40% of total agricultural sector and >12% of GDP. This contribution would have been much greater had the animal by-products been also efficiently utilized. Efficient utilization of by-products has direct impact on the economy and environmental pollution of the country. Non-utilization or under utilization of by-products not only lead to loss of potential revenues but also lead to the added and increasing cost of disposal of these products. Non-utilization of animal by-products in a proper way may create major aesthetic and catastrophic health problems. Besides pollution and hazard aspects, in many cases meat, poultry and fish processing wastes have a potential for recycling raw materials or for conversion into useful products of higher value. Traditions, culture and religion are often important when a meat by-product is being utilized for food. Regulatory requirements are also important because many countries restrict the use of meat by-products for reasons of food safety and quality. By-products such as blood, liver, lung, kidney, brains, spleen and tripe has good nutritive value. Medicinal and pharmaceutical uses of by-product are also highlighted in this review. Waste products from the poultry processing and egg production industries must be efficiently dealt with as the growth of these industries depends largely on waste management. Treated fish waste has found many applications among with which the most important are animal feed, biodiesel/biogas, dietectic products (chitosan), natural pigments (after extraction) and cosmetics (collagen). Available information pertaining to the utilization of by-products and waste materials from meat, poultry and fish and their processing industries has been reviewed here.

683 citations

Journal ArticleDOI
TL;DR: The fundamentals of microbial kinetics and continuous culture models are presented and the effect of temperature and inhibitors on the intrinsic kinetic rates is discussed, and Stoichiometric and bioenergetic considerations are reviewed.
Abstract: The fundamentals of microbial kinetics and continuous culture models are presented. The kinetics of the anaerobic treatment processes are reviewed recognizing that anaerobic degradation of complex, polymeric organic materials is a combination of series and parallel reactions. Such reactions include hydrolysis, fermentation, anaerobic oxidation of fatty acids, and methanogenesis. The intrinsic rates of each step are reviewed and literature data summarized. Whenever possible, available kinetic information is summarized on the basis of substrate composition (such as carbohydrates, proteins, and lipids). The effect of temperature and inhibitors on the intrinsic kinetic rates is discussed. Stoichiometric and bioenergetic considerations are reviewed. Mass transfer limitations (both external and internal) associated with biofilms and microbial agglomerates, in general, and their effect on the intrinsic kinetic rates are presented. Areas requiring further research are identified.

638 citations