• 제목/요약/키워드: Protease production

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Application Potential of Hurdle Technology by Combination of Bacteriocin Produced by Lactobacillus brevis DK25 and Potassium Benzoate (Lactobacillus brevis DK25의 박테리오신과 안식향산칼륨과의 혼용에 의한 Hurdle Technology 적용 가능성)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.364-374
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    • 2011
  • Lactobacillus brevis DK25 isolated from Dongchimi was identified by physiological and biochemical tests and 16S rDNA sequence analysis. Bacteriocin of L. brevis DK25 exhibits inhibitory activity against Enterococcus faecalis and Listeria monocytogenes when using agar well diffusion method. Maximal production of bacteriocin was reached in the beginning of the stationary phase, and inhibitory activity declined after the late stationary phase. This result suggested that bacteriocin was produced in a growth-associated manner. Complete inactivation of bacteriocin activity was observed after treatment with protease, but the activity was stable between pH 4-9 and heat resistant (30 min at $100^{\circ}C$). Bacteriocin showed a concentration-dependent antimicrobial activity against L. monocytogenes KCTC 3569. Moreover, the application experiment showed that combination of bacteriocin (320 AU/ml) with potassium benzoate (0.05%) could significantly reduce the counts of L. monocytogenes KCTC 3569 in mayonnaise during storage at 4 or $25^{\circ}C$ for 10 days. Thus, bacteriocin from L. brevis DK25 may be used for hurdle technology by combination with potassium benzoate in order to increase pathogenic bacteria inactivation in food processing and food safety control.

Fermentation of Black Garlic Wine and its Characteristics (흑마늘 와인의 발효 및 그 특성)

  • Ha, Sung Min;Choi, Hye Jung;Shin, Gyeong Yeon;Ryu, Beung Ho;Joo, Woo Hong
    • Journal of Life Science
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    • v.26 no.7
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    • pp.796-804
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    • 2016
  • In the present study, we screened suitable yeasts for wine fermentation and evaluated the fermentative characteristics of Saccharomyces sp. BCNU 6006 and its anti-oxidant activities. Firstly, various yeasts were isolated from Makgeolli, fruits, and fermented foods. Then, the preliminary selections of suitable yeasts were made using an enzymatic activity assay of glucosidase, glycosidase, protease and tolerance to ethanol and SO2. In addition, the production of biogenic amines and hydrogen sulfide was also monitored. The 9 yeast strains initially selected were determined to belong to the genera Saccharomyces and Kazachtania phylogenetically. We investigated the optimal conditions for wine fermented with black garlic juice (BGJ). The optimal conditions of alcohol fermentation using BGJ were 26 brix, 28℃, and 10 days. Finally, the fermentation products of black garlic wine (BGW) fermented with Saccharomyces sp. BCNU 6006 exhibited 15.03% ethanol, 12 brix of sugar, and pH 4.01. The contents of total polyphenol, total flavonoid, tannin, and 5-HMF compound of BGW were 3.85 mg/ml, 0.51mg/ml, 5.90 mg/ml, and 0.07 mg/ml respectively, lower than that of BGJ. DPPH radical scavenging activity, ABTS radical scavenging activity, and reducing power of BGW were 90.77%, 95.20% and 1.261 respectively, lower than that of BGJ. Superoxide anion (O2-) radical scavenging activity was 94.42%, higher than that of BGJ. Based on the above results, the industrial potential of Saccharomyces sp. BCNU 6006 as a wine-making yeast was confirmed in the present study.

Effect of Red Pepper Varieties on the Microflora, Enzyme Activities and Taste Components of Traditional Kochujang during Fermentation (고추품종을 달리한 전통식 고추장의 숙성중 미생물, 효소활성 및 맛성분의 변화)

  • Shin, Dong-Hwa;Kim, Dong-Han;Choi, Ung;Lim, Mi-Sun;An, Eun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1050-1057
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    • 1997
  • Microbial counts, enzyme activities and taste components of traditional kochujangs prepared with the powders from 4 different varieties of red pepper, were investigated during 90 days fermentation for the industrial production of traditional kochujang. The viable cell counts of anaerobic bacteria in kochujangs did not change remarkably during fermentation, however, aerobic bacterial counts showed a rapid increase up to 90 days of aging. The yeasts in all kochujang samples increased until 60 days of aging and than decreased. After 90 days of aging, the count of aerobic bacteria in Kumtop kochujang was higher than those of others. The activities of liquefying amylase decreased during the aging, but those of saccharogenic amylase increased at 60 days of aging. The activities of neutral protease were higher than those of acidic protease, and increased during the middle and last period of aging. The major free sugars of kochujang were maltose and glucose, and their contents were higher in Hongkwang kochujang. The major organic acids of kochujang were succinic, formic and citric acid, followed by lactic acid. Succinic acid content in kochujang decreased during fermentation, whereas formic and citirc acids were increased. The major free amino acids were serine, proline, glutamic acid, aspartic acid and alanine. Kumtop kochujang contained the highest amount of total free amino acids. Among the nucleotides and related components in kochujang, cytidine-5-monophosphate was the most abundant component at the begining of aging period, while hypoxanthine increased remarkably during fermentation. Hanwang kochujang was higher in the content of nucleotides than others. Capsaicin contents of kochujang decreased during aging and those of Jangter kochujang was higher than that of others. Sensory evaluation showed that Jangter kochujang was significantly better than Kumtop kochujang in overall acceptability, but there were no appreciable differences in color and flavor.

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β-Xylosidase and β-mannosidase in combination improved growth performance and altered microbial profiles in weanling pigs fed a corn-soybean meal-based diet

  • Liu, Shaoshuai;Ma, Chang;Liu, Ling;Ning, Dong;Liu, Yajing;Dong, Bing
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1734-1744
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    • 2019
  • Objective: In this study, two glycosidases (XMosidases), ${\beta}$-xylosidase and ${\beta}$-mannosidase, were investigated on their in vitro hydrolysis activities of feed and on the improvement of growth performance in vivo in weanling pigs. Methods: Enzyme activities of XMosidases in vitro were evaluated in test tubes and simulation of gastric and small intestinal digestion, respectively, in the presence of NSPase. In vivo study was performed in 108 weaned piglets in a 28-d treatment. Pigs were allotted to one of three dietary treatments with six replicate pens in each treatment. The three treatment groups were as follows: i) Control (basal diet); ii) CE (basal diets+CE); iii) CE-Xmosidases (basal diets+ CE+${\beta}$-xylosidase at 800 U/kg and ${\beta}$-mannosidase at 40 U/kg). CE was complex enzymes (amylase, protease, xylanase, and mannanase). Results: In vitro XMosidases displayed significant activities on hydrolysis of corn and soybean meal in the presence of non-starch polysaccharide degrading enzymes (xylanase and ${\beta}$-mannanase). In vitro simulation of gastric and small intestinal digestion by XMosidases showed XMosidases achieved $67.89%{\pm}0.22%$ of dry matter digestibility and $63.12%{\pm}0.21%$ of energy digestibility at $40^{\circ}C$ for 5 hrs. In weanling pigs, additional XMosidases to CE in feed improved average daily gain, feed conversion rate (p<0.05), and apparent total tract digestibility of crude protein (p = 0.01) and dry matter (p = 0.02). XMosidases also altered the gut bacterial diversity and composition by increasing the proportion of beneficial bacteria. Conclusion: Addition of a complex enzyme supplementation (contained xylanase, ${\beta}$-mannanase, protease and amylase), XMosidases (${\beta}$-xylosidase and ${\beta}$-mannosidase) can further improve the growth performance and nutrient digestion of young pigs.

A Study of the Diversity and Profile for Extracellular Enzyme Production of Aerobically Cultured Bacteria in the Gut of Muraenesox cinereus (갯장어(Muraenesox cinereus) 장으로부터 호기적 조건에서 분리된 미생물의 다양성 및 세포외 효소 생산능 분석에 관한 연구)

  • Lee, Yong-Jik;Oh, Do-Kyoung;Kim, Hye Won;Nam, Gae-Won;Sohn, Jae Hak;Lee, Han-Seung;Shin, Kee-Sun;Lee, Sang-Jae
    • Journal of Life Science
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    • v.29 no.2
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    • pp.248-255
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    • 2019
  • This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.

Studies on the Brewing of Kochuzang (Red Pepper Paste) by the Addition of Yeasts (효모첨가(酵母添加)에 의(依)한 고추장의 양조(釀造)에 관(關)한 연구(硏究))

  • Lee, Taik-Soo
    • Applied Biological Chemistry
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    • v.22 no.2
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    • pp.65-90
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    • 1979
  • This study was conducted to establish the brewing method which would be useful for the production of Kochuzang. Kojis, which were made from various materials and microorganisms under a covered condition, were investigated and compared. Yeasts (Saccharomyces rouxii and Torulopsis versatilis) were added to Kochuzang, and the enzyme activity, microflora, chemical composition, nitrogen content, alcohol content and free sugars of Kochuzang were investigated and analysed. The results obtained are as follows: 1. Koji making (1) Glutinous rice-soybean group was superior to glutinous rice group in the saccharogenic and liquefying amylase activities of three day-Koji. (2) Protease activity (acid, neutral and alkaline) of glutinous rice-soybean Koji, which was inoculated with Aspergillus oryzae A, was increased till the 5th day, while other groups showed maximum activity after the 3rd day. (3) The maximum cellulose activity of Aspergillus oryzae B-Koji and A-Koji was observed after the 2nd day and the 3rd day, respectively. High cellulose activity of Aspergillus oryzae B-Koji and A-Koji was respectively shown in glutinous rice group and glutinous rice-soybean group at maximum. (4) Compared with glutinous rice Koji, glutinous rice-soybean Koji gave larger number of yeast and aerobic bacteria. 2. Kochuzang Fermentation (1) Each Kochuzang group shoved different liquefying and saccharogenic amylase activities. The highest activities were generally shown in 10 to 40 days after mashing and remarkably reduced in the last stage of aging. (2) Protease activities of each group were strong in order of acid, neutral and alkaline protease. Especially acid protease showed highest activity at the 40th to 50th day Kochuzang. (3) Each group showed maximum cellulase activity in the 40th and 50th day-Kochuzang and then decreased. (4) Osmophilic yeast of yeast-added Kochuzang after one-month aging was distinctively outnumbered compared with non-yeast-added Kochuzang, but two groups were similar after two months. (5) Yeast-added group and non-added group gave almost the same number of halophilic lactic acid bacteria in Kochuzang, but the non-added group gave slightly larger number of aerobic bacteria than the yeast-added group. (6) Amino nitrogen contents in all test group were increased rapidly till the 60th day of Kochuzang aged. After that the contents were increased slowly. (7) Ethyl alcohol contents of 20day-fermented Kochuzang were high in order of Saccharomyces rouxii-added group, Torulopsis versatilis-added group, Saccharomyces rouxii and Torulopsis versatilis mixed group and non-yeast-added group. But all test group showed about 2% in ethyl alcohol content after 40days of aging. (8) Alcohol content in the 7 month-aged Kochuzang of all test groups was high in order of ethyl alcohol, n-butyl alcohol, n-propyl alcohol and iso-propyl alcohol. Torulopsis versatilis-added group had the highest value of ethyl alcohol, n-propyl alcohol and n-butyl alcohol. (9) Reducing sugar in Kochuzang was increased after 20 days of aging compared with the 10days-ferment. The reducing sugar content in Saccharomyces rouxii-added group was distinctively small compared with that of other groups, decreasing after 30days of aging. (10) Rhamnose, fructose, glucose and maltose were isolated from the 10 day fermented Kochuzang. Raffinose was also found after 300 days-aged group, and fructose content was high in the 300days-aged Kochuzang. However, glucose content was smaller than that of 10days-fermented Kochuzang. (11) For the organoleptic tests of Kochuzang, taste, flavour and color of yeast-added group were superior to the non yeast-added group. Especially the complex yeast group among the yeast added groups were the best of all. Yeast-added group after 300 days of aging took higher paint in flavour test than that of non-added group. Therefore, brewing method like complex yeast added group seems to be advantageous for short time brewing Kochuzang.

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Properties of Plywood Bonded with Adhesive Resins Formulated with Enzymatically-Hydrolyzed Rapeseed Flour (유채박의 효소 가수분해물로 조제한 접착제를 사용한 합판의 접착특성)

  • Yang, In;Han, Gyu-Seong;Choi, In-Gyu;Kim, Yong-Hyun;Ahn, Sye-Hee;Oh, Sei-Chang
    • Journal of the Korean Wood Science and Technology
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    • v.40 no.3
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    • pp.164-176
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    • 2012
  • In the present study, rapeseed flour (RSF), which is a by-product from the production of edible oil and biodiesel extracted from rapeseed, was used to develop alternative adhesives for the production of plywood panels. To examine the effects of the enzyme on the adhesive properties and formaldehyde emission of the RSF-based adhesive resins, three enzymes, such as cellulase (CEL), pectinase (PEC) and protease (ALC), were used either separately or together. As a crosslinking agent, PF prepolymers, which were prepared with 1.5, 1.8 and 2.1 mole formaldehyde and 1 mol phenol (1.8-, 2.1- and 2.4-PF), were added into the RSF hydrolyzates. The adhesive resins formulated with CEL- or CEL-PEC-RSF hydrolyzates and 1.8-F/P PF prepolymers exhibited excellent adhesive strengths and formaldehyde emission. The tensile shear strength and formaldehyde emission of the plywood panels bonded with the formulate resins were satisfied with the minimum requirement of the KS standard for ordinary plywood panels (0.6 N/$mm^2$). In addition, formaldehyde emissions of the plywood panels approached to that of E0 specified in the KS standard (0.5 mg/${\ell}$), and even had much better than those of commercial UF glue mixes. Overall, the use of RSF-based adhesive resins for the production of plywood panels might provide durable adhesive properties and an environmentally friendly substitute for petroleum-based adhesive resins. However, further researches - the increase of solid content of RSF-based adhesives for reducing press time and the microscopic observation of plywood specimen for identifying the relationship between tensile shear strength and the penetration of adhesives into wood structure - are required to commercialize the RSF-based adhesives.

Effects of Spider-Derived Protease (Arazyme®) Supplementation of Corn-Soy Diets on the Performance in Laying Hens at the Late Production (산란 후기 사료 내 거미(Nephila clavata) 유래 단백질 분해효소제(Arazyme®)의 첨가 급여가 난 생산성에 미치는 영향)

  • Kim, J.U.;Kim, J.Y.;Kim, J.S.;Lee, B.K.;Lee, S.Y.;Lee, W.S.;You, S.J.;Ahn, B.K.;Kim, E.J.;Park, H.Y.;Son, K.H.;Shin, D.H.;Kang, C.W.
    • Korean Journal of Poultry Science
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    • v.36 no.1
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    • pp.47-55
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    • 2009
  • This study was carried out to investigate the effects of spider-derived protease [Arazyme (AZ)] supplementation on egg production, egg quality, cecal microflora and viscosity of intestinal contents in laying hens at the late production. A total of two hundred, 59-wks-old, Hy-Line Brown layers were divided into four groups, placed into 5 replicates per group (50 birds per group) and fed one of four diets containing 0.05, 0.075 or 0.1% AZ or control for 5 wks. The results showed that egg weight in group fed the diet containing 0.05% AZ was significantly higher than those of other groups (P<0.05). There were no significant differences in egg interior and eggshell qualities among the groups. The serum parameters such as total cholesterol, the levels of GOT and GPT were not influenced by AZ feeding. There were no significant differences in the cecal number of total microbes, Coliforms and lactic acid bacteria among the groups. The viscosity and ammonia concentration of intestinal contents in the groups fed AZ were significantly reduced compared to those of the control (P<0.05). In conclusion, laying hens fed AZ supplemented diet improved egg weight as compared with that of control. In addition, the AZ supplemented diet reduced the viscosity and ammonia concentration of intestinal contents, reflecting a significant feed enzyme effect.

Isolation of High Yielding Alkaline Protease Mutants of Vibrio metschnikovii Strain RH530 and Detergency Properties of Enzyme

  • Chung, So-Sun;Shin, Yong-Uk;Kim, Hee-Jin;Jin, Ghee-Hong;Rho, Hyune-Mo;Lee, Hyune-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.349-354
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    • 2000
  • Abstract A facultative alkalophilic gram-negative Vibrio metschnikovii strain RH530, isolated from the wastewater, produced several alkaline proteases (VAP) including six alkaline serine proteases and a metalloprotease. From this strain, high yielding YAP mutants were isolated by NTG treatment. The isolated mutant KS1 showed nine times more activity than the wild-type after optimization of the culture media. The production was regulated by catabolite repression when glucose was added to the medium. The effects of several organic nitrogen sources on the production of the YAP were investigated to avoid catabolite repression. The combination of 4% wheat gluten meal (WGM), 1.5% cotton seed flour (eSF), and 5% soybean meal (SBM) resulted in the best production when supplemented with 1% NaCl. The YAP showed a resistance to surfactants such as $sodium-{\alpha}-olefin$ sulfonate (AOS), polyoxy ethylene oxide (POE), and sodium dodecyl sulfate (SDS), yet not to linear alkylbenzene sulfonate (LAS). However, the activity of the YAP was restored completely when incubated with LAS in the presence of POE or $Na_2SO_4$. The YAP was stable in a liquid laundry detergent containing 6.6% SLES (sodium lauryl ether sulfate), 6.6% LAS, 19.8% POE, and stabilizing agents for more than two weeks at $40^{\circ}C$, but the stability was sharply decreased even after 1 day when incubated at $60^{\circ}C$. A washing performance test with the YAP exhibited it to be a good washing power by showing 51 % and 60% activity at $25^{\circ}C{\;}and{\;}40^{\circ}C$, respectively, thereby indicating that the YAP also has a good detergency at a low temperature. All the results suggest that the YAP produced from the mutant strain KSI has suitable properties for use in laundry detergents.rgents.

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Relationship between production of exoenzymes and serotypes of Pseudomonas aeruginosa isolated from clinical specimens and hospital environments (녹농균(綠膿菌)의 균체외효소산생능(菌體外酵素産生能)과 그의 혈청형(血淸型)과의 관계(關係))

  • Moon, Hong-Yong;Cho, Yag-Ja
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.47-54
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    • 1980
  • Exoenzymes, protease(P) and elastase(E) produced by Pseudomonas aeruginosa are reported to have close relationship with pathogenicity of Pseudomonas aeruginosa. Productibility of exoenzymes P and E were studied and compared in environmental isolates from hospital environments and clinical isolates from various clinical specimens, also, the relationship between their enzyme production and serotype were reviewed. 1. Clinical isolates were typed into nine serotypes A, B, C, D, E, F, G, H and I. Serotype E had the highest incidence of 24%, followed by B with 16.8%, G, 15.1% and C, 9.3%. 2. Environmental isolates were, typed as serotype B, C, E, F, G, H, I, K and M. Serotype I had the highest incidence of 26.6%, followed by C, F and M each incidence of 14.3%. 3. In the typing of the above two groups, serotypes A and D were found only in the clinical isolates and serotypes K, and M were found only in the environmental isolates. Serotypes J and L were found in neither clinical isolates nor environmental isolates. 4. In the distribution of serotypes from various clinical specimens, serotype G among isolates from pus showed incidence of 20.4%, and serotypes E and B were 19.5% separately. Serotype E had incidence of 22.6% and 20.0% in urine and sputa respectively, showing a high rate compared to the other serotypes. 5. The incidence of strains producing both exoenzymes P and E was 77.8% in the preserved strains of clinical isolates and 76.2% in the environmental isolates. There were no significant difference between the two groups. 6. Serotypes A and H, which are preserved strains from clinical isolates showed productibility of both exoenzymes P and E, the other serotypes showed productibility of various combination of exoenzymes. Among the environmental isoaltes, production of both exoenzymes P and E were seen in serotypes E, F, G, H, I and K and no serotype produced only P or E. 7. In ability to produce exoenzymes of isolates from sources of various clinical specimens, strains producing both exoenzymes P and E were found most frequently in pus with incidence rate of 82.0%, followed by 80.0% in sputum and urine. 8. Almost all the fresh strains of clinical isolates were producers of both exoenzymes P and E.

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