scholarly journals OPTIMASI JENIS DAN KADAR SUMBER NITROGEN SERTA pH MEDIUM UNTUK PRODUKSI PROTEASE DARI ISOLAT HTcUM6.2.2 DARI TAUCO SURABAYA

2017 ◽  
Vol 2 (2) ◽  
pp. 98 ◽  
Author(s):  
Monika Rahayu ◽  
Evi Susanti
Keyword(s):  
Nitrogen Source ◽  
Protease Activity ◽  
Skim Milk ◽  
Protease Production ◽  
Production Medium ◽  
Medium Ph ◽  
Ph Optimum ◽  
Research Stage ◽  
Nitrogen Percentage

AbstrakPenelitian ini bertujuan untuk  mengetahui sumber nitrogen, kadar sumber nitrogen dan pH optimum untuk produksi protease isolat HTcUM6.2.2. Penelitian ini merupakan penelitian eksperimental laboratoris menggunakan Rancangan Acak Lengkap (RAL) satu faktorial masing-masing yaitu jenis, kadar sumber nitrogen, serta pH medium dengan tahap penelitian terdiri dari peremajaan, validasi kemurnian isolat HTcUM6.2.2, produksi ekstrak kasar protease isolat HTcUM6.2.2,  uji aktivitas protease. Hasil penelitian menunjukkan sumber nitrogen optimum untuk menghasilkan protease dari isolat HTcUM6.2.2 adalah susu skim dan limbah cair tahu. Kadar limbah cair tahu optimum untuk produksi protease dari isolat HTcUM6.2.2 sebesar 10 %. Produksi protease cukup tinggi dan  relatif konstan antara pH 6 sampai 8.  Aktivitas protease tertinggi sebesar 0,817 U/ml dicapai dengan penggunaan 10 % limbah cair tahu pada medium produksi, pH = 7 selama 3 hari. Kata kunci: protease, tauco, isolat, nitrogen, limbah cair tahu   AbstractThis study aims to determine the optimum of the nitrogen source, percentage of nitrogen source and pH for the production of protease from isolate HTcUM6.2.2. This research is a laboratory experimental with a research stage comprising inoculation and validation of the purity of the isolates HTcUM6.2.2, production of crude extract of protease to determine source of nitrogen, percentage of nitrogen source and pH optimum, determination of protease activity. The results showed that the optimum source of nitrogen to produce proteases from the HTcUM6.2.2 isolate was skim milk and tofu wastewater. Percentage of nitrogen source optimum to produce protease of  HTcUM6.2.2 isolate was 10 % of tofu wastewater. Protease production is relatively high and constantance at pH 6 to 8. The highest protease activity was achieved by the use of 10 % tofu wastewater at production medium, pH = 7 for 3 days was 0,817 U/mL. The keywords: protease, tauco, isolate, nitrogen, tofu wastewater

Production and characterisation of thermostable alkaline protease from Bacillus subtilis isolated from LA hot spring, Terengganu

2021 ◽  
Vol 16 (7) ◽  
pp. 84-91
Author(s):  
Maslinda Alias ◽  
Hakim Che Harun Mohammad ◽  
Ashraf Razali Nurul ◽  
Jasnizat Saidin ◽  
Nazaitulshila Rasit ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Alkaline Protease ◽  
Protease Activity ◽  
Hot Spring ◽  
Skim Milk ◽  
Industrial Applications ◽  
Protease Production ◽  
Significant Activity ◽  
Original Activity ◽  
Optimum Ph

This research aims to produce thermostable alkaline protease from Bacillus subtilis isolated from La Hot Spring, Terengganu, Malaysia. The study was also conducted to determine the optimum conditions for protease production and stability by considering several parameters including pH, temperature and salt concentration. All seven bacteria were screened on skim milk agar overnight at 37 °C. Three strains with the highest proteolytic activity were identified in protease specific medium. The thermostable alkaline protease had an optimum temperature of 60 °C which achieved 85.73, 82.90 and 83.05 U/mL of protease activity for the three strains respectively. Furthermore, the strains exhibited significant activity of more than 90% from their original activity. Meanwhile, the optimum pH for protease production was pH 9 with the protease activity of 76.76, 79.71 and 88.39 U/mL for TB4, TB6 and TB9 strains, respectively. Proteases were found stable at pH 9 where the loss did not exceed 30% of its original activity. Collectively, all of the data emphasised that proteases from B. subtilis were alkaline thermostable proteases in accordance with a recent report. The finding highlights the viability of the proteases for biotechnological and industrial applications.

scholarly journals Production of alkaline protease from Aspergillus oryzae isolated from seashore of Bay of Bengal

2018 ◽  
Vol 10 (4) ◽  
pp. 1210-1215 ◽  
Author(s):  
Raghu Ram M ◽  
Suman Kumar Yepuru
Keyword(s):  
Aspergillus Oryzae ◽  
Wheat Bran ◽  
Bay Of Bengal ◽  
Alkaline Protease ◽  
Protease Activity ◽  
Skim Milk ◽  
Protease Production ◽  
Chemical Parameters ◽  
Oil Cakes ◽  
Physical And Chemical

Aspergillus oryzae isolatedon  Potato dextrose agar  from soil samples of kottakoduru seashore of Bay of Bengal, Andhra Pradesh, India seashore of Bay of Bengal by spread plate method and was screened for alkaline protease production on Skim milk containing agar plates and identified by clear zones of protein hydrolysis around colonies. Different physical and chemical parameters such as pH, temperature, substrate concentration and incubation time were optimized for the better production of alkaline protease. The maximum protease activity was found at pH of 8 containing 10% wheat bran at 300C, after 72 hours of fermentation.ZnSO4was effective activator for protease activity and sodium dsulphate had shownmore than 50% inhibition of enzyme activity. Among the different oil cakes used for the production of enzyme the Sesame  oil cake proved to be suitable substrate after wheat bran for the production of protease by Aspergillus oryzae.

scholarly journals CULTIVATION CONDITIONS FOR PROTEASE PRODUCTION BY A THERMO-HALOSTABLE BACTERIAL ISOLATE PLS A

2019 ◽  
Vol 19 (1) ◽  
pp. 18-23
Author(s):  
Teuku M. Iqbalsyah ◽  
Malahayati Malahayati ◽  
Atikah Atikah ◽  
Febriani Febriani
Keyword(s):  
Protease Activity ◽  
Hot Springs ◽  
Skim Milk ◽  
Dry Weight ◽  
Protease Production ◽  
Cultivation Conditions ◽  
Thermostable Enzymes ◽  
Cultivation Period ◽  
Protease Enzyme ◽  
Nacl Concentration

Polyextremophiles have increasingly been utilised to produce thermostable enzymes with better stability in multiple extreme conditions. This study reports the screening results of four new bacterial isolates (PLS A, PLS 75, PLS 76 and PLS 80), isolated from an under water hot springs, in producing thermo-halostable protease enzyme. Optimum cultivation conditions for the protease production were also studied. Screening of protease-producing isolates was conducted using Thermus solid medium enriched with 3% skim milk and 0.5% casein. The growth of the isolates showing protease activity was monitored by measuring the cell dry weight and protease activity during 24 h cultivation period. The activity was also measured at various cultivation conditions, i.e. temperature, pH and salt concentrations. Amongst the four isolates, only PLS A showed the ability to produce protease. The optimum cultivation conditions for protease production were observed at 65°C, pH 7 for 18 h incubation. The activity increased with the addition of 1% NaCl concentration (0.085 Unit/mL). The ability of PLS A isolate to produce thermo-halostable protease was encouraging as they could potentially be used in industries requiring the enzyme with multiple extremes. 

scholarly journals Enzymatic activity of alkaline protease from Bacillus cereus TD5B and its application as sheep skin dehairing agent

2021 ◽  
Vol 21 (2) ◽  
pp. 105-118
Author(s):  
Nanung Agus FITRIYANTO ◽  
MUSTHOFIYAH MUSTHOFIYAH ◽  
MUHLISIN MUHLISIN ◽  
Ambar PERTIWININGRUM ◽  
Novita KURNIAWATI ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Bacillus Cereus ◽  
Alkaline Protease ◽  
Protease Activity ◽  
Skim Milk ◽  
Protease Production ◽  
Activity Measurement ◽  
Randomized Design ◽  
Chrome Tanning ◽  
Agar Media

This study aims to determine the enzymatic activity of extracellular alkaline protease from Bacillus cereus TD5B and its potential application as a sheep skin dehairing agent. The B. cereus TD5B was screened for extracellular alkaline protease production on skim milk agar media, while its alkaline protease activity and the application were measured at 1%, 1.5%, and 2%. The application of alkaline protease from B. cereus TD5B as a sheep skin dehairing agent was observed through histological examination and physical properties measurement after chrome-tanning with lime and Na2S as control. The study was conducted in a completely randomized design, and the quantitative data were analyzed using Duncan’s Multiple Range Test. The results showed that a clear zone was seen surrounding B. cereus, indicating the bacteria’s proteolytic activity. The protease activity measurement showed that 2% of alkaline protease had the highest enzymatic activity at 144.75 U/mL/min. The highest tensile strength of sheep leather was obtained after dehairing at 1% alkaline protease concentration (350.26 kg/cm2), even though the highest elongation was obtained at 2% (34.92%). In contrast, different concentrations showed similar shrinkage temperatures at 90°C. This study concludes that the optimum alkaline protease concentration from Bacillus cereus TD5B as a sheep dehairing agent was 2%.

Casein, Fluorescein Isothiocyanate as a Substrate for Measuring Protease Production by Psychrotrophic Bacteria

1987 ◽  
Vol 50 (9) ◽  
pp. 744-749 ◽  
Author(s):  
GENEVIEVE L. CHRISTEN ◽  
M. SENICA
Keyword(s):  
Protease Activity ◽  
Free Amino ◽  
Skim Milk ◽  
Fluorescein Isothiocyanate ◽  
Enzyme Concentration ◽  
Protease Production ◽  
Amino Groups ◽  
Bacterial Populations ◽  
Psychrotrophic Bacteria ◽  
Bacterial Protease

Fluorescein isothiocyanate-labelled casein was evaluated as a substrate for measuring bacterial protease activity in milk. Using protease from Bacillus amyloliquefaciens, effects of substrate type, pH, temperature and enzyme concentration were determined. The procedure was sensitive to .012 unit of enzyme activity. The procedure was then used as a method to detect protease production by psychrotrophic bacteria growing in pasteurized skim milk at 7°C for 5 d. Three psychrotrophic bacteria were added at three different initial populations. Bacterial population, protease activity and cumulative free amino groups were monitored daily. Bacterial populations were not correlated with either protease activity or cumulative free amino groups. Pro-tease activity and cumulative free amino groups were correlated (r=.46, p=.001). Both protease activity and cumulative free amino groups peaked as the microorganisms entered the late log phase (approximately 108 colony forming units per ml). This occurred after incubation for 2 d at 7°C in two replications and after 3 d in the third. Both values decreased with continued storage.

Cytotoxic Effect of Pseudomonas aurogenosa Protease on Cancer Cells

2019 ◽  
Vol 10 (03) ◽  
Author(s):  
Ghanyia J. Shanyoor ◽  
Fatima R. Abdul ◽  
Nehad A. Taher ◽  
Ihsan A. Raheem
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Cytotoxic Effect ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) and the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method and it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column and the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , and one normal cell line Ref ( Rat embryonic fibroblast ) . The cancer and normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8and 0.16 mg/ml) then incubated for additional 48h at 37C0 and the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andslight toxicity ( 37.12% ) on normal cell line (Ref) in a concentration (0.8mg/ml).

Constraints in Adoption ofModern Farm Machines by Tribal Farmers in Ramgarh District of Jharkhand

2019 ◽  
Vol 6 (03) ◽  
Author(s):  
PK SUNDARAM ◽  
BIKASH SARKAR ◽  
UJJWAL KUMAR ◽  
AP ANURAG ◽  
DK RAGHAV ◽  
...  
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Tribal Farmers ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) andamp; the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method andamp; it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column andamp; the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , andamp; one normal cell line Ref ( Rat embryonic fibroblast ). The cancer andamp; normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8andamp; 0.16 mg/ml) then incubated for additional 48h at 37C 0 andamp; the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andamp;slight toxicity ( 37.12% )on normal cell line (Ref) in a concentration (0.8mg/ml).

Polarographic study of hydrolysis of [8-lysine]vasopressin and its derivatives with blood serum of pregnant women

1980 ◽  
Vol 45 (4) ◽  
pp. 1099-1108 ◽  
Author(s):  
Mikuláš Chavko ◽  
Michal Bartík ◽  
Evžen Kasafírek
Keyword(s):  
Blood Serum ◽  
Pregnant Women ◽  
Degree Of Hydrolysis ◽  
Polarographic Study ◽  
Ph Optimum ◽  
Lysine Vasopressin ◽  
Rate Of Hydrolysis ◽  
Hydrolysis Of ◽  
Vasopressin Analogues

A polarographic study of the hydrolysis of [8-lysine]vasopressin and some hormonogens of the vasopressin series with the blood serum of women in the last week of pregnancy was studied. The dependence of hydrolysis on pH (pH optimum: 7.4-7.50, substrate concentration (Km 1.2 . 10-5M), pH stability and thermal stability were determined. The rate of hydrolysis of individual vasopressin analogues decreases in the order: [8-lysine]vasopressin > Nα-glycyl-prolyl[8-lysine]-vasopressin > Nα-leucyl-[8-lysine]vasopressin > Nα-alanyl-[8-lysine]vasopressin > Nα-phenyl alanyl-[8-lysine]vasopressin > Nα-diglycyl-[8-lysine]vasopressin > Nα-prolyl-[8-lysine]vasopressin > Nα-triglycyl-[8-lysine]vasopressin > Nα-sarcosyl-glycyl-[8-lysine]vasopressin. The degree of hydrolysis gradually increases to a multiple with the length of the pregnancy in consequence of the presence of oxytocine. However, vasopressin is also hydrolysed to a small extent with the enzymes from the blood sera of non-pregnant women. Under similar analytical conditions oxytocin was not hydrolysed with the sera of non-pregnant women and therefore oxytocin is a more suitable substrate than vasopressin for polarographic determination of serum oxytocinase.

Determination of Bisphenol A in Milk and Dairy Products by High-Performance Liquid Chromatography with Fluorescence Detection

2003 ◽  
Vol 66 (8) ◽  
pp. 1439-1443 ◽  
Author(s):  
JEONG-HUN KANG ◽  
FUSAO KONDO
Keyword(s):  
Bisphenol A ◽  
Dairy Products ◽  
High Performance ◽  
Solid Phase ◽  
Skim Milk ◽  
The European Union ◽  
Condensed Milk ◽  
Oasis Hlb ◽  
Milk And Dairy Products

This study was conducted to develop a selective and sensitive method for the determination of bisphenol A (BPA) levels in milk and dairy products. A method based on solvent extraction with acetonitrile and solid-phase extraction (SPE) was developed for the analysis of BPA in milk, yogurt, cream, butter, pudding, condensed milk, and flavored milk, and a method using two SPE cartridges (OASIS HLB and Florisil cartridge) for skim milk was also developed. The developed methods showed good recovery levels (77 to 102%) together with low detection limits (1 μg/liter for milk, yogurt, pudding, condensed milk, flavored milk, and skim milk and 3 μg/liter for cream and butter). These methods are simple, sensitive, and suitable for the analysis of BPA in milk and dairy products. When 40 milk and dairy products were analyzed by the proposed methods, BPA was not identified in noncanned products, but its levels ranged from 21 to 43 μg/kg in canned products, levels that were 60- to 140-fold lower than the migration limits in the European Union and Japan.

570. A medium for the simultaneous enumeration and preliminary identification of milk microflora

1955 ◽  
Vol 22 (1) ◽  
pp. 43-47 ◽  
Author(s):  
Kathleen O. Donovan ◽  
J. M. Vincent
Keyword(s):  
Acid Production ◽  
Skim Milk ◽  
Biochemical Properties ◽  
Viable Count ◽  
Hydrogen Ion ◽  
Ion Concentration ◽  
Hydrogen Ion Concentration ◽  
Pasteurized Milk ◽  
Preliminary Identification

A medium has been developed that permits the viable count of milk bacteria to be combined with the determination of biochemical properties likely to be important in milk itself. This has involved the modification of standard glucose-tryptone skim-milk agar by incorporation of two indicators to detect alkali as well as acid production, substitution of lactose for glucose, and increasing the quantity of skim milk for the detection of proteolysis and casein precipitation. The medium has proved particularly valuable in the study of the thermoduric flora of pasteurized milk. The phenomenon of casein precipitation is, however, less reliably determined than are changes in hydrogen-ion concentration and proteolysis.

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