• Journal of Life Science
• /
• v.31 no.8
• /
• pp.761-770
• /
• 2021

In this study, the optimal medium composition for enhancing protease production was established by the Bacillus strain isolated from Makgeolli, a traditional fermented food, using the response surface methodology. B. amyloliquefaciens SRCM115785 was selected as the protease producer by productivity analysis and identified by 16S rRNA gene sequencing. Plackett-Burman design (PBD) was introduced to analyze the effect of each component on protease production among the 11 selected medium components. As a result, glucose, yeast extract, and beef extract were finally selected as factors for enhancing protease production. Central composite design (CCD) analysis was designed as a method to determine the optimal concentration of each component for protease production and the concentration of each medium composition for maximum protease production was predicted to glucose 6.75 g/l, yeast extract 12.42 g/l and beef extract 17.48 g/l. The suitability of the experimental model was proved using ANOVA analysis and as a result of quantitative analysis to prove this, the amount of increase was 230.47% compared to the LB medium used as a control. Through this study, the optimization of medium composition for enhancing protease production was established, and based on this, it is expected that it can be efficient use of protease as an industrial enzyme.

• Journal of Life Science
• /
• v.21 no.2
• /
• pp.309-315
• /
• 2011

Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.

• Journal of the Korean Wood Science and Technology
• /
• v.32 no.5
• /
• pp.39-44
• /
• 2004

This study was carried out to investigate the tenderizing effect of Neungi mushroom (Sarcodon aspratus) powder and its protease. The addition of Neungi mushroom powder and its protease enhanced water retention values (WRY) of meat. The WRY of meat was increased 26.8% by protease addition, compared to 13.8% WRV by sugar addition. This increase in WRY derived to the increase of water soluble fraction in the meat texture by hydrolysis of meat protein, and had the meat tenderized. Concerned to the meat tenderizing effect, the addition of Neungi mushroom powder and its protease have decreased of meat hardness and gave similar tenderizing effect, as compared to commercial tenderizer, papain. The decreasing rates of meat hardness were 51.6% of Neungi mushroom powder, 58.5% of its protease, and 563% of commercial tenderizer, papain. This tenderizing effect of protease attributed to the degradation of muscle fiber protein in meat, such as actin, myosin and connectin etc. The addition of Neungi mushroom to foods gives significant changes in food color, mainly decreasing lightness.

• Journal of Animal Science and Technology
• /
• v.66 no.2
• /
• pp.326-339
• /
• 2024

This study was conducted to investigate the effects of protease supplementation and different nutrient density of diets in growing-finishing pigs. A total of one hundred-eight crossbred growing pigs ([Landrace × Yorkshire] × Duroc) with an initial body weight (BW; 18.74 ± 3.46 kg) were used for 15 weeks. Pigs were randomly assigned to six dietary treatments with 6 replicates of 3 pigs per pen in a 3 × 2 factorial through the following arrangement: Three groups of protease (1, Basal diets; 2, Protease A: 125 mg/kg protease derived from Streptomyces sps; 3, Protease B: 100 mg/kg protease derived from Bacillus licheniformis) at two different nutrient density diets (1, Basal requirement; 2, 0.94%-0.98% higher than requirement in dietary protein and 50 kcal/kg in energy). High nutrient (HN) diets showed higher average daily gain (ADG) (p < 0.05) and apparent total tract digestibility (ATTD) of crude protein (CP) (p < .0001) compared to basal nutrient (BN) diets during growing periods. Supplementation of protease showed higher BW (p < 0.05) and ADG (p < 0.05) compared to non-supplementation of protease during growing periods. Also, supplementation of protease showed higher ATTD of CP (p < 0.01), ATTD of gross energy (p < 0.05) and decreased blood urea nitrogen (BUN) level (p = 0.001) compared to non-supplementation of protease during finishing periods. Pigs which fed the protease showed decreased ammonia (NH₃) emissions (p < 0.05) during experiment periods and decreased hydrogen sulfide (H₂S) emissions (p < 0.01) during finishing periods. Interactions between nutrient density and protease were observed, which decreased the feed conversion ratio (p < 0.05) in HN diets without protease compared to BN diets without protease during weeks 4 to 6. Also, interaction between nutrient density and protease was observed, which resulted in improved ATTD of CP (p < 0.01) in response to PTA supplementation with HN diets during the finishing period. In conclusion, supplementation of protease reduces NH₃ in feces and BUN in whole blood by increasing the digestibility of CP and improves growth performance. Also, diets with high nutrient density improved growth performance and nutrient digestibility in growing periods.

• KSBB Journal
• /
• v.5 no.4
• /
• pp.411-414
• /
• 1990

The extraction behavior of an alkaline protease using reversed micelles was investgated. The reversed micellar solution consisted of AOT in isooctane. It was found that distribution of arkaline protease into the organic phase increased at lower pH, lower ionic strength, and higher AOT concentration. When the real fermentation broth was extracted of alkaline protease, an activity yield of 20% and a purification factor of 2.0 were obtained.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.5
• /
• pp.677-684
• /
• 2000

Pseudomonas sp. BK7, an alkalophile, displayed the highest growth and protease activity when grown in a fermenter which was controlled at a pH level of 9.0, and the enzyme production was significantly enhanced by the increase of agitation speed. Two forms of alkaline proteases (BK7-1 and BK7-2) were fractionated and purified to near homogeneity. Protease BK7-1 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B, DEAE-Sepharose, and Sephadex G-75 column chromatographies. The molecular weights of proteases BK7-1 and BK7-2 determined by gel filtration chromatography were 20,700 and 40,800, respectively. The $K_m$ value, isoelectric point, and optimum pH of protease BK7-1 were 2.55 mg/ml, 11.0, and 11.0, respectively, whereas those of protease BK7-2 were 1.57 mg/ml, 7.2, and 10.0, respectively. Both proteases were practically stable in the pH range of 5-11. The optimum temperatures for the activities of both protease BK7-1 and BK7-2 were $50^{\circ}C$ and $45^{\circ}C$, respectively. About 56% of the original protease BK7-2 activity remained after being treated at $50^{\circ}C$ for 30 min but protease BK7-1 was rapidly inactivated at above $25^{\circ}C$. Both proteases were completely inhibited by phenylmethane sulfonyl fluoride, a serine protease inhibitor. Protease BK7-2 was stable against EDTA, EGTA, STP, and detergents such as SDS and LAS, whereas protease BK7-1 was found to be unstable.

• Journal of Life Science
• /
• v.7 no.3
• /
• pp.228-233
• /
• 1997

A thiol protease has been isolated and partially purified from encysted brine shrimp Artemia franciscana using a four-step procedure(filtration, salting out, gel filtration and ion exchange chromatography). The optimum pH of the enzyme for caseinolytic activity was appeared to be 3.0, and the enzymematic activity was stable up to pH 6.0 but lost completely at the pH higher than 8.0. The optimal temperature of the enzyme was appeared to be 35$^{\circ}$C, and ninety percent of the enzyme activity was lost at 45$^{\circ}$C. Various metal ions, e.g., zinc, copper, iron, inhibited the enzyme activity; however, heavy metal chelator, e.g., EDTA, stimulated the enzyme activity. The protease was concluded to be a member of the thiol group protease, since it was inhibited by thiol protease inhibitors and iodoacetate. The protease was also concluded to be a acid protease based on optimum pH.

• Journal of the Korean Society of Clothing and Textiles
• /
• v.22 no.2
• /
• pp.224-232
• /
• 1998

Protease is mixtured in detergent to remove protein-soil easily. It must not act on the any fiber except protein-soil during laundry. So the purpose of this study is to investigate how protease is affect the fiber, particulary the protein-fiber. For this purpose, silk, wool and nylon are selected as samples, and the extent of the damage was estimated as tensile strength and surface condition (that is fibrillation). The results are as follows. The tensile strength of fiber treated with protease were lowered at enzyme concentration 0.1%, temperature 4$0^{\circ}C$ , and, as washing time was longer, it was lowered more. And it was showed that the surface of fibers were fiblliated by protease during washing. From this results, it was found that protease damaged protein-fiber. The damage of silk was the largest of all, and wool was less damaged than silk, because it has the scale (cuticle) on the outside. Additionary, an influence of surfactant on damage of fiber was little about three fibers, but, the fibers were damaged more by the binary nonionic-surfactant and protease mixture than by protease only.

• Microbiology and Biotechnology Letters
• /
• v.19 no.5
• /
• pp.450-455
• /
• 1991

A protease hyperproducer, ampicillin resistant mutant, Serratia sp. SMNT-1 was selected from Serratia marcescens ATCC 21074 by mutagenesis. The protease productivity of this strain was about 11 times as much as that of the parental strain. The enzyme showed maximal activity at pH 9.0 and $40^{\circ}C$ and was stable over the pH range from 6.0 to 10.0 at $4^{\circ}C$, whereas it was unstable at $37^{\circ}C$ in alkaline condition. the enzyme was inactivated by heating at $60^{\circ}C$ for 10 min. The enzyme was inactivated by EDTA and reactivated by $Zn^{2+}, Co^{2+},\; and \; Mn^{2+}$, but the proteoiytic activity of the enzyme was not affected by DFP. From the above results, the protease produced by Serratia sp. SMNT-1 was classified as a metalloprotese.

• Biomolecules & Therapeutics
• /
• v.2 no.2
• /
• pp.155-160
• /
• 1994

Human immunodeficiency virus type 1 (HIV-1) protease is essential for the replication of the virus and it is therefore an attractive target for antiviral drugs of HIV-1. Several dipeptide isosteres containing 2-isoxazoline or $\alpha$-hydroxy ketomethylene have been synthesized and their inhibitory effects on the HIV-1 protease examined. The enzymatically active HIV-1 protease was purified to homogeniety from E. coli transformed with a recombinant plasmid (pMAL-pro) containing the entire gene encoding the protease. The purified protease had the substrate specificity with Km value of 9.8$\mu$M when an undecapeptide His-Lys-Ala-Arg-Val-Leu-(p-nitro)Phe-Glu-Ala-Nle-Ser-amide was used as a substrate, and the products from the substrate after specific cleavage by HIV-1 protease were analyzed by HPLC. The synthetic compounds containing dipeptide isosteres showed specific inhibitory effects while a dipeptide isostere containing an isoxazoline ring inhibited the HIV-1 protease competitively with Ki value of 500 $\mu$M. Even if the inhibition effects of HIV-1 protease were not very high, these novel dipeptide isosteres can be used as key structural moieties for developing specific inhibitors of HIV-1 protease.