@article{wang_borwornpinyo_shih_2007, title={Sup35NM-His6 aggregates: A prion-like protein useful in prion degradation studies}, volume={40}, ISSN={["0141-0229"]}, DOI={10.1016/j.enzmictec.2006.09.019}, abstractNote={A prion-like protein, Sup35NM-His6, which is safe, easy to produce and able to aggregate into stable amyloid, was developed. This study was to test the feasibility of using Sup35NM-His6 amyloid as a marker for standard sterilization methods known to be effective in inactivating infectious prions. Also Sup35NM-His6 aggregates were spiked into cow brain tissue homogenates to mimic prions in tissues to determine if the aggregate protein could be specifically detected by Western blot at the nanogram level. Like mammalian prions, the proteinase K (PK) resistant fractions of Sup35NM-His6 remain intact after autoclaving. Treatments with 0.1N NaOH or 1.0% NaOCl also resulted in PK undigestible residues. Exposure to the strong denaturant guanidine thiocyanate (>3 M) destabilized and made the protein PK-digestible. Under strong alkaline conditions of 1.0N NaOH, 2.5% NaOCl, or a combination of NaOH (0.1 and 1.0N) with autoclaving, Sup35NM-His6 was completely hydrolyzed such that it was no longer detectable by Western blot. Overall, these tests suggest that Sup35NM-His6 could be a useful tool to assess the effectiveness of prion degradation for the prevention of TSE.}, number={4}, journal={ENZYME AND MICROBIAL TECHNOLOGY}, author={Wang, Jeng-Jie and Borwornpinyo, Rattana and Shih, Jason C. H.}, year={2007}, month={Mar}, pages={976–981} }
 @article{shih_wang_2006, title={Keratinase technology: from feather degradation and feed additive,to prion destruction}, volume={1}, ISBN={1749-8848}, DOI={10.1079/pavsnnr20061042}, abstractNote={Abstract}, number={042}, journal={CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources}, author={Shih, J. C. H. and Wang, J. J.}, year={2006}, pages={6} }
 @article{wang_greenhut_shih_2005, title={Development of an asporogenic Bacillus licheniformis for the production of keratinase}, volume={98}, ISSN={["1364-5072"]}, DOI={10.1111/j.1365-2672.2004.02515.x}, abstractNote={Aims:  Bacillus licheniformis PWD‐1 is a keratin‐degrading, spore‐forming bacterium isolated from a poultry waste digester. A sporulation‐deficient mutant of B. licheniformis PWD‐1, named B. licheniformis WBG, was developed and characterized.}, number={3}, journal={JOURNAL OF APPLIED MICROBIOLOGY}, author={Wang, JJ and Greenhut, WB and Shih, JCH}, year={2005}, pages={761–767} }
 @article{kang_wang_shih_lanier_2004, title={Extracellular production of a functional soy cystatin by Bacillus subtilis}, volume={52}, ISSN={["1520-5118"]}, DOI={10.1021/jf049711x}, abstractNote={A recombinant Bacillus subtilis producing soy cystatin was developed by subcloning with a soy cystatin gene cloned in Escherichia coli. An active form of cystatin against the cysteine protease from Pacific whiting fillets contaminated with Myxosporidia parasite was constitutively expressed and secreted extracelluarly into the medium. Two gene fragments of signal peptides from kerA and sacB were introduced and compared for secretion efficiency of cystatin. The secretion level of active cystatin improved with the signal peptide of kerA when compared to that of sacB. Inhibitor activity was reduced rapidly after peak expression of the target protein at 36 h of fermentation. The addition of 1% glucose, a suppressor of protease, into the medium sustained the increase of the cystatin activity during fermentation. This study introduced a potential new method for fermentation production of cystatin.}, number={16}, journal={JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY}, author={Kang, IS and Wang, JJ and Shih, JCH and Lanier, TC}, year={2004}, month={Aug}, pages={5052–5056} }
 @article{wang_rojanatavorn_shih_2004, title={Increased production of Bacillus keratinase by chromosomal integration of multiple copies of the kerA gene}, volume={87}, ISSN={["0006-3592"]}, DOI={10.1002/bit.20145}, abstractNote={Abstract}, number={4}, journal={BIOTECHNOLOGY AND BIOENGINEERING}, author={Wang, JJ and Rojanatavorn, K and Shih, JCH}, year={2004}, month={Aug}, pages={459–464} }
 @article{wang_swaisgood_shih_2003, title={Bioimmobilization of keratinase using Bacillus subtilis and Escherichia coli systems}, volume={81}, ISSN={["0006-3592"]}, DOI={10.1002/bit.10485}, abstractNote={Abstract}, number={4}, journal={BIOTECHNOLOGY AND BIOENGINEERING}, author={Wang, JJ and Swaisgood, HE and Shih, JCH}, year={2003}, month={Feb}, pages={421–429} }
 @article{odetallah_wang_garlich_shih_2003, title={Keratinase in starter diets improves growth of broiler chicks}, volume={82}, ISSN={["1525-3171"]}, DOI={10.1093/ps/82.4.664}, abstractNote={The objective of this study was to determine the efficacy of a broad-spectrum protease enzyme, PWD-1 keratinase, upon supplementation to corn-soy starter diets on growth performance of broiler chickens. Three experiments were conducted. In each experiment, 1-d-old broiler chicks were randomly assigned to 24 cage pens of eight birds per pen in a completely randomized design of five experimental treatments and grown to 21 or 26 d of age. Treatments in experiments 1 and 2 were control (C, 21.39% CP), low protein (LP, 18% CP), and LP supplemented with 0.05, 0.1, or 0.15% enzyme preparation (wt/wt). Treatments in experiment 3 were control (C), C+ 0.1% enzyme preparation (C+E) fed starting at either 1 or 5 d of age, LP and LP+ 0.1% enzyme preparation (LP+E). Feeding the LP+E diet produced numerically higher BW at 21 d of age (experiments 1 and 3) and a significantly higher BW at 26 d of age (experiment 2; 1,025 and 1,032 g vs. 965 g for 0.1 and 0.15% vs. LP, respectively, P < 0.05). Feed conversion ratio (FCR) was also improved when chicks were fed the LP+E diet both at 21 (experiment 3) and 26 d of age (experiment 2). In experiment 3, supplementing the C diets with 0.10% enzyme resulted in improvements (P < 0.05) in BW whether the enzyme was supplemented starting at 1 d (767 vs. 695 g for C+E vs. C, respectively) or 5 d of age (764 vs. 695 g for C+E vs. C, respectively). FCR was numerically improved. Furthermore, diets supplemented with the enzyme at any level resulted in reduction of jejunal viscosity at 22 and 27 d of age (P < 0.05). Results of these experiments indicate that the growth of broiler chickens can be significantly improved by dietary supplementation with PWD-1 keratinase.}, number={4}, journal={POULTRY SCIENCE}, author={Odetallah, NH and Wang, JJ and Garlich, JD and Shih, JCH}, year={2003}, month={Apr}, pages={664–670} }
 @article{wang_swaisgood_shih_2003, title={Production and characterization of bio-immobilized keratinase in proteolysis and keratinolysis}, volume={32}, ISSN={["0141-0229"]}, DOI={10.1016/S0141-0229(03)00060-7}, abstractNote={Extracellular production of keratinase–streptavidin fusion protein (KER–STP) was accomplished by the cloning of Bacillus subtilis with a transforming plasmid carrying the kerA-stp fusion gene. The fusion protein was readily immobilized onto a biotinylated solid matrix by mixing in the culture medium. The properties and reaction kinetics of free and immobilized keratinase (KE) were characterized and compared. Heat stability and pH tolerance were greatly improved by immobilization, but the catalytic efficiency (kcat/Km) was reduced by eightfold. The yield of bio-immobilization using bioselective adsorption of the fusion protein was approximately 20%, as estimated from the activity of free KE. HPLC analysis of reaction products demonstrated the hydrolysis of feather keratin, casein, and bovine serum albumin (BSA) by the immobilized KE. The rates of reactions are lower than those of the free enzyme. On the other hand, the stability of the enzyme was greatly improved.}, number={7}, journal={ENZYME AND MICROBIAL TECHNOLOGY}, author={Wang, JJ and Swaisgood, HE and Shih, JCH}, year={2003}, month={Jun}, pages={812–819} }
 @article{wang_shih_1999, title={Fermentation production of keratinase from Bacillus licheniformis PWD-1 and a recombinant B-subtilis FDB-29}, volume={22}, ISSN={["1367-5435"]}, DOI={10.1038/sj.jim.2900667}, abstractNote={Fermentation scale-up was studied for the production of keratinase by Bacillus licheniformis PWD-1, the parent strain, and B. subtilis FDB-29, a recombinant strain. In both strains, keratinase was induced by proteinaceous media, and repressed by carbohydrates. A seed culture of B. licheniformis PWD-1 at early age, 6-10 h, is crucial to keratinase production during fermentation, but B. subtilis FDB-29 is insensitive to the seed culture age. During the batch fermentation by both strains, the pH changed from 7.0 to 8.5 while the keratinase activity and productivity stayed at high levels. Control of pH, therefore, is not necessary. The temperature for maximum keratinase production is 37 degrees C for both strains, though B. licheniformis is thermophilic and grows best at 50 degrees C. Optimal levels of dissolved oxygen are 10% and 20% for B. licheniformis and B. subtilis respectively. A scale-up procedure using constant temperature at 37 degrees C was adopted for B. subtilis. On the other hand, a temperature-shift procedure by which an 8-h fermentation at 50 degrees C for growth followed by a shift to 37 degrees C for enzyme production was used for B. licheniformis to shorten the fermentation time and increase enzyme productivity. Production of keratinase by B. licheniformis increased by ten-fold following this new procedure. After respective optimization of fermentation conditions, keratinase production by B. licheniformis PWD-1 is approximately 40% higher than that by B. subtilis FDB-29.}, number={6}, journal={JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY}, author={Wang, JJ and Shih, JCH}, year={1999}, month={Jun}, pages={608–616} }