• 한국식품영양학회지
• /
• 제7권1호
• /
• pp.51-57
• /
• 1994

A nuluk, a Korean traditional koji for brewing, was made with wheat flour and Aspergillus usamii mot. shirousamii S1 which had strong abilities in producing amylase and protease. The cultural conditions for the production of saccharogenic and proteolytic enzymes were tested. The productivities of saccharogenic and dextrogenic enzymes were improved when nuluk was made with unsteamed wheat flour as compared with steamed one, but those of proteolytic enzyme and organic acid were reduced. The addition of water containing 0.5% of hydrochloric acid was unfavorable for the production of saccharogenic, dextrogenic and proteolytic enzymes. The optimum ratios of water added to wheat flour for the production of saccharogenic enzyme and proteolytic enzyme were 32% and 28%, respectively on the basis of wheat flour. The optimum temperatures for the production of saccharogenic enzyme and proteolytic enzyme were 36$^{\circ}C$ and 28$^{\circ}C$, respectively. The activity of saccharogenic enzyme reached its maximum after 120 hours of cultivation at 36$^{\circ}C$, but that of proteolytic enzyme 96 hours. The productivity of saccharogenic enzyme was enhanced when the nuluk was molded after 24 hours of precultivation but that of proteolytic enzyme was reduced as compared with no molding.

• Journal of Microbiology and Biotechnology
• /
• 제20권5호
• /
• pp.950-957
• /
• 2010

Serratiopeptidase (SRP), a 50 kDa metalloprotease produced from Serratia marcescens species, is a drug with potent anti-inflammatory property. In this study, a powerful statistical design, evolutionary operation (EVOP), was applied to optimize the media composition for SRP production in shake-flask culture of Serratia marcescens NRRL B-23112. Initially, factors such as inoculum size, initial pH, carbon source, and organic nitrogen source were optimized using one factor at a time. The most significant medium components affecting the production of SRP were identified as maltose, soybean meal, and $K_2HPO_4$. The SRP so produced was not found to be dependent on whey protein, but rather was notably induced by most of the organic nitrogen sources used in the study and free from other concomitant protease contaminant, as revealed by protease inhibition study. In addition, experiments were performed using different sets of EVOP design with each factor varied at three levels. The experimental data were analyzed with a standard set of statistical formula. The EVOP-optimized medium, with maltose 4.5%, soybean meal 6.5%, $K_2HPO_4$ 0.8%, and NaCl 0.5% (w/v), gave a SRP production of 7,333 EU/ml, which was 17-fold higher than the unoptimized media. The application of EVOP resulted in significant enhancement of SRP production.

• Applied Biological Chemistry
• /
• 제2권
• /
• pp.17-21
• /
• 1961

The manufacture of riboflavine fortified Dwen-Jang has been tried with an exceedingly riboflavine productive Aspergillus oryzae #612 mutant which has been developed by the authors. Both the rice and barley koji of this mutant and Aspergillus sojae have been prepared. Their riboflavine production, saccharifying and protease activities have begin compared The riboflavine fortified Dwen-Jang has been manufactured using the barley koji of riboflavine productive mutant. Their riboflavine content and qualities have been studied comparing with an ordinary Dwen-Jang which has been prepared with the barley kojo of A. sojae strain. The following results have been obtained. (1) The baley koji was superior in riboflavine production and protease activity, inferior in saccharifying ability than rice koji both with A. oryzae #612 and A. sojae. (2) In barley koji, the mutant, A. oryzae #612, produces 1.5 times riboflavine than A. sojae and shows stronger saccharifying and protease activities than the latter. (3) The riboflavine fortified Dwen-Jaug manufactured contained $5.2{\gamma}/g$ of riboflavine, about 1.5 times that of A. sojae. The higher contents of free sugar and free amjno nitrogen have been observed than the ordinary Dwen-Jang manufactured with A. sojae.

• 한국균학회지
• /
• 제24권4호통권79호
• /
• pp.329-336
• /
• 1996

From five years' previous work, the fungal isolates of Scopulariopsis were reported to be important flora at the late stage of meju fermentation. Mainly, the white or pale brown powders of spore mass of these fungi were observed on the surfaces of rectangular mejus, and to be an important sign for well-done Korean traditional home - made mejus. Out of the five isolates previously collected and stored, two kinds of Scopulariopsis isolates were identified as S. brevicauli and S. fusca. The microscopic differences between two were found to be branching patterns of annellophore and ornamentations of spore wall (warty and smooth). However, the intermediate form between two ornamentations of spore wall were also observed in our isolates. This observation was consistent with other result made from the protein electrophoresis. The isolates of Scopulariopsis were considered to be similar or superior to those of Aspergillus species, as compared with production of protease and amylase related enzymes. Thus, these isolates were speculated to be important fungi in Korean traditional home - made meju fermentation and also in production of protease and amylase.

• 한국미생물·생명공학회지
• /
• 제48권4호
• /
• pp.515-524
• /
• 2020

With the increasing demand for enzymes in industrial applications there is a growing need to easily produce industrially important microbial enzymes. This study was carried out to screen the indigenous refinery bacterial isolates for their production of two industrially important enzymes i.e. lipase and protease. A total of 15 bacterial strains were isolated using Soil Extract Agar media from the oil-contaminated environment and one was shown to produce high quality lipase and protease enzymes. The culture conditions (culture duration, temperature, source of nitrogen, carbon, and pH) were optimized to produce the optimum amount of both the lipase (37.6 ± 0.2 Uml-1) and the protease (41 ± 0.4 Uml-1) from this isolate. Productivity of both enzymes was shown to be maximized at pH 7.5 in a medium containing yeast extract and peptone as nitrogen sources and sucrose and galactose as carbon sources when incubated at 35 ± 1℃ for 48 h. Bacterial strain SAB06 was identified as Bacillus licheniformis (MT250345) based on biochemical, morphological, and molecular characteristics. Further studies are required to evaluate and optimize the purification and characterization of these enzymes before they can be recommended for industrial or environmental applications.

• 한국식품과학회지
• /
• 제9권1호
• /
• pp.31-35
• /
• 1977

Rhizopus japonicus S-62가 생산(生産)하는 acid protease에 관(關)한 연구(硏究)로서, 효소(酵素)의 생산조건(生産條件) 및 조효소(粗酵素)의 특성(特性)을 검토(檢討)하여 다음과 같은 결과(結果)를 얻었다. 1. 밀기울에 배양(培養)하는 경우 최적배양(最適培養) 시간(時間)은 48시간(時間) 첨수량(添水量)은 $100{\sim}120%$이었다. 2. 각종(各種) 배지중(培地中) 밀기울 배지(培地)가 가장 우수(優秀)하였다. 3. 밀기울에 sucrose, fructose, $NH_4Cl$, $NaH_2PO_4$의 첨가(添加)는 protease생산(生産)을 크게 증가(增加)시켰다. 4.본효소(本酵素)의 작용최적(作用最適)pH는 $2.4{\sim}2.6$, Opt. temp.는 약(約) $40^{\circ}C$, 안정(安定) pH 범위(範圍)는 $2.5{\sim}5.0$이고, $40^{\circ}C$이하(以下)에서는 안정(安定)하나 그 이상(以上)의 온도(溫度)에서는 급격(急激)하게 불활성화(不活性化)되었다.

• Mycobiology
• /
• 제33권1호
• /
• pp.23-29
• /
• 2005

The effects of different concentrations of three amino acids as carbon and or nitrogen sources on mycelial dry weights, changes in pH values of synthetic medium, ammonia secretion and extracellular protease activity by three zoosporic fungi, pathogens of fish and shellfish, were studied. As compared with the control, the addition of isoleucine and aspartic acid as nitrogen sources were generally stimulative for mycelial dry weight production whereas phenylalanine was inhibitory irrespective to the tested fungal species. When amino acids served as carbon and nitrogen sources, the mycelial dry weights of the three fungi were increased (mostly non-significantly) relative to untreated control but weights were decreased as the concentrations of the three amino acids raised. The addition of individual amino acids as carbon and nitrogen sources to the medium significantly increased pH values of the medium comparable to the control. The addition of each of the three amino acids as carbon and nitrogen sources to the medium significantly induced ammonia secretion by the three species of zoosporic fungi. Ammonia secretion in synthetic medium amended with amino acids as nitrogen source raised by the three zoosporic fungi relative to untreated control except in case of Achlya racemosa treated with isoleucine. Extracellular protease activity was almost promoted in case of Achlya proliferoides and Saprolegnia furcata cultures treated with isoleucine and aspartic acid individually in presence of glucose and vice versa in case of phenylalanine. However, extracellular protease activity of A. racemosa decreased compared with the control at various concentrations of isoleucine and both phenylalanine and aspartic acid assumed inconsistent effects. Extracellular protease activity of the three zoosporic fungi in the medium devoid of glucose varied depending upon zoosporic fungal species, the tested amino acid and the applied concentrations. The values of protease activity were approximately less two folds than that obtained in presence of glucose.

• 한국식품영양과학회지
• /
• 제18권3호
• /
• pp.348-355
• /
• 1989

Alkaline protease 생성능이 강한 Aspergillus fumigatus 균주를 토양에서 분리하고, 생성효소를 정제하여 특성을 조사한 결과 최적 pH는 9.0, pH안정성은 $pH\;8.0{\sim}10.0$, 최적온도는 $50^{\circ}C$였으며, $50^{\circ}C$이하의 온도에서 안정하나 그 이상의 온도에서는 급격한 효소 불활성화를 보였고, 금속염 $Mn^{++},\;Cu^{++},\;Ba^{++},\;Mg^{++}$ 등에 의해서 활성이 다소 증대되나 $K^+,\;Fe^{+++},\;Ag^{++},\;Pb^{++},\;Na^+,\;Ca^{++},\;Hg^+,\;Zn^{++}$에 의해 저해를 받았다. 활성저해제인 EDTA, 2,4-DNP, ${\varepsilon}-amino$ caproic acid에는 큰 저해를 받지 않으나, PCMB에 많은 저해를 받는 것으로 미루어 활성 부위가 SH기인 cystein protease로 추정되었다. Km값은 $8.33{\times}10^{-4}mole/{\ell}$, Vmax는 $47.62{\mu}g/min$였으며, casein과 hemoglobin을 trypsin보다 더 잘 분해하고, casein을 hemoglobin보다 잘 분해하였다.

• Journal of Animal Science and Technology
• /
• 제62권6호
• /
• pp.840-853
• /
• 2020

The present study was conducted to investigate the effect of a multi-protease on production indicators of broiler chickens fed a crude protein and amino acid deficient-diets for 35 days immediately after hatch. A total of 448 one-day-old Ross 308 male broiler chicks were allocated in a completely randomized design into one of eight dietary treatments (positive control [PC], negative control [NC: minus 0.5% from PC, and minus 2% of lysine, methionine, threonine and methionine plus cysteine], extreme negative control [ENC: minus 1% from PC, minus 4% of lysine, methionine, threonine and methionine plus cysteine], and plus multi-protease 150 or 300 g per ton [e. g., PC-150]; PC, PC-150, NC, NC-150, NC-300, ENC, ENC-150, ENC-300) to give eight replicates with seven birds in a battery cage. Body weight, average daily gain, average daily feed intake, feed conversion ratio, and mortality were measured every week. Carcass traits, proximate analysis of breast meat, and ileum digestibility were analyzed on day 21 and 35. Feeding a multi-protease (i.e., more than 150 g/ton) for 35 days immediately after hatching improved feed efficiency and ileum digestibility (i.e., dry matter, crude protein, and energy) compared to their counterparts (i.e., diets without multi-protease: PC, NC, and ENC). In conclusion, our results indicated that broiler chickens fed nutrients deficient-diet (i.e., crude protein and amino acids) supplemented a multi-protease had an ability to compensate and (or) improve their growth performance commensurate with increased ileal digestibility for 35 days immediately after hatch.

• Animal Bioscience
• /
• 제36권7호
• /
• pp.1034-1043
• /
• 2023

Objective: Two serine protease inhibitors, peptidase inhibitor 3 (PI3) and secretory leukocyte protease inhibitor (SLPI), play important roles in protease inhibition and antimicrobial activity, but their expression, regulation, and function at the maternal-fetal interface in pigs are not fully understood. Therefore, we determined the expression and regulation of PI3 and SLPI in the endometrium throughout the estrous cycle and at the maternal-fetal interface in pigs. Methods: Endometrial tissues during the estrous cycle and pregnancy, conceptus tissues during early pregnancy, and chorioallantoic tissues during mid to late pregnancy were obtained, and the expression of PI3 and SLPI was analyzed. The effects of the steroid hormones estradiol-17β (E2) and progesterone (P4) on the expression of PI3 and SLPI were determined in endometrial explant cultures. Results: PI3 and SLPI were expressed in the endometrium during the estrous cycle and pregnancy, with higher levels during mid to late pregnancy than during the estrous cycle and early pregnancy. Early-stage conceptuses and chorioallantoic tissues during mid to late pregnancy also expressed PI3 and SLPI. PI3 protein and SLPI mRNA were primarily localized to endometrial epithelia. In endometrial explant cultures, the expression of PI3 was induced by increasing doses of P4, and the expression of SLPI was induced by increasing doses of E2 and P4. Conclusion: These results suggest that the PI3 and SLPI expressed in the endometrium and conceptus tissues play an important role in antimicrobial activity for fetal protection against potential pathogens and in blocking protease actions to allow epitheliochorial placenta formation.