Isolation and characterization of novel and efficient protease producing bacteria from drinking water resources

2019 ◽  
Vol 20 (1) ◽  
pp. 157-164
Author(s):  
S. Jeevan Chandra ◽  
A. Shiva Shanker ◽  
Pavan Kumar Pindi

Abstract An attempt has been made to explore the stability of protease enzyme (isolated from Bacillus sp.) by statistical method. More than 100 isolates were screened for extracellular protease activity derived from various potable water samples of Mahabubnagar district, Telangana State, India. The activity of protease is found to be varying from sample to sample, the highest being reported by the isolate from water sample of Kalwakurthy mandal, Mahabubnagar district and therefore was selected for further studies. The 16S rRNA (ribosomal ribonucleic acid) gene sequence of the isolate showed closest similarity with Bacillus sp. and the sequence was submitted to National Center for Biotechnology Information (NCBI) gene bank (accession number GU566359) and the culture was deposited in three international culture deposition centers (KCTC-13725: MTCC-10465: JCM-16713). The present study revealed that, this Bacillus sp. showed a greater amount of protease production with the inherent characters of thermo, alkali and oxidant stability which makes it a potential alternative protease producing strain in various industrial applications.

2018 ◽  
Vol 10 (1) ◽  
pp. 8 ◽  
Author(s):  
Rachmat Rizaldi ◽  
Woro Hastuti Setyantini ◽  
Sudarno Sudarno

AbstrakLamun adalah tumbuhan sejati yang hidup di perairan pantai yang kurang dimanfaatkan dalam bidang perikanan, selain sebagai bioindikator kualitas air laut. Beberapa mikroorganisme yang berasosiasi dengan lamun Enhalus acoroides antara lain benthos, kapang, bakteri dan plankton. Bakteri proteolitik merupakan bakteri yang mampu menghasilkan enzim protease. Enzim perotese merupakan enzim yang paling banyak digunakan dalam kehidupan. Bakteri merupakan sumber enzim yang paling banyak digunakan dibandingkan dengan tanaman dan hewan. Tujuan penelitian ini adalah untuk mengetahui jenis isolat bakteri proteolitik yang berasosiasi dengan lamun Enhalus acoroides di Taman Nasional Baluran, Situbondo. Penelitian ini menggunakan metode survei dengan dianalisis secara deskriptif dengan bantuan tabel dan gambar. Hasil yang peroleh menunjukkan bahwa terdapat 12 isolat bakteri yang berasosisasi dengan lamun Enhalus acoroides. Terdapat empat isolat yang tergolong sebagai bakteri proteolitik karena mampu mendagradasi kasein dalam media TSA + 2% NaCL yang ditambah 1 % susu skim, tampak dari pembentukan zona bening. Pengamatan morfologi koloni dan sel serta pengujian biokimia dari keempat isolat (EA-1, EA-2, EA-9 dan EA-10) terdapat kesamaan karakteristik dengan empat genus bakteri berturut-turut yaitu Staphylococcus sp., Plesiomonas shigeloides, Bacillus sp., Pseudomonas sp.AbstractSeagrass is a true living plants underutilized coastal waters in the field of fisheries, as well as bio-indicators of the quality of sea water. Some microorganisms associated with seagrass Enhalus acoroides among others benthos, fungi, bacteria and plankton. Proteolytic bacteria are bacteria that are capable of producing the enzyme protease. Protease enzyme is an enzyme that is most widely used in life. Bacteria are a source of enzymes that are most widely used compared to plants and animals. The purpose of this study was to determine the type of proteolytic bacterial isolates associated with seagrass Enhalus acoroides in Baluran National Park, Situbondo. This study used survey method with descriptive analysis with tables and figures. The results obtained that there are 12 bacterial isolates associated with seagrass Enhalus acoroides. There are four isolates were classified as proteolytic bacteria because it can degrade casein in TSA media + 2 % NaCL plus 1% skim milk which is evidenced by the formation of clear zones. Observations colony morphology and cells, as well as testing of Biochemistry of the four isolates (EA-1, EA-2, EA-9 and EA-10), were obtained, with similar characteristics to the four genera of bacteria in a row as follows Staphylococcus sp., Plesiomonas shigeloides, Bacillus sp., Pseudomonas sp. 


2007 ◽  
Vol 50 (2) ◽  
pp. 217-223 ◽  
Author(s):  
Eliana Tiemi Ito ◽  
Geni Varéa-Pereira ◽  
Dalva Tomoe Miyagui ◽  
Maria Helena Pimenta Pinotti ◽  
Pedro Manoel Oliveira Janeiro Neves

Studies were carried out on extracellular protease production by Beauveria bassiana CG432 in liquid medium containing glucose and yeast extract. B. Bassiana presented active growth after lag period of 24 h., produced 80% of the total of the extracellular protease activity in 48 h which was maximum on the 5th culture day. The extracellular protease presented optimum activity at 60ºC, was stable up to 1M Cl-, maintained the stability during 15 day at 4ºC and -18ºC, but was not stable if frozen repeatedly.


2008 ◽  
Vol 57 (5) ◽  
pp. 639-645 ◽  
Author(s):  
J. P. Chenel ◽  
R. D. Tyagi ◽  
R. Y. Surampalli

The volume of sludge produced annually is very high and poses serious disposal problems. The traditional methods of sludge disposal produce secondary pollutants. Therefore, the alternate or suitable solution is reuse of sludge in an ecofriendly approach. Biotechnology is an interesting tool to add value to the processes involved in wastewater and wastewater sludge disposal/reuse. In this context, a study was carried out on thermophilic bacterial strains that produce thermostable proteases. The bacterial strains were first isolated from municipal wastewater sludge. In contrast to the conventional strains used in industries, like Bacillus sp., the new strains were Gram-Negative type. In semi-synthetic medium, a maximal protease activity of 5.25 IU/ml (International Unit per ml) was obtained at a pH of 8.2 and at a temperature of 60 °C, which is higher than the stability temperature of 37 °C for a similar protease obtained from the conventional producer Bacillus licheniformis. Moreover, growth and protease activity of the strains were tested in wastewater sludge. It is expected that the complexity of sludge could stimulate/enhance the protease production and their characteristics. In conclusion, reuse of wastewater sludge will help to reduce their quantity as well as the value-added products produced will replace chemical products used in industries.


Author(s):  
Ghanyia J. Shanyoor ◽  
Fatima R. Abdul ◽  
Nehad A. Taher ◽  
Ihsan A. Raheem

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).


2019 ◽  
Vol 6 (03) ◽  
Author(s):  
PK SUNDARAM ◽  
BIKASH SARKAR ◽  
UJJWAL KUMAR ◽  
AP ANURAG ◽  
DK RAGHAV ◽  
...  

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).


Mathematics ◽  
2020 ◽  
Vol 8 (9) ◽  
pp. 1434 ◽  
Author(s):  
Wonhee Kim ◽  
Sangmin Suh

For several decades, disturbance observers (DOs) have been widely utilized to enhance tracking performance by reducing external disturbances in different industrial applications. However, although a DO is a verified control structure, a conventional DO does not guarantee stability. This paper proposes a stability-guaranteed design method, while maintaining the DO structure. The proposed design method uses a linear matrix inequality (LMI)-based H∞ control because the LMI-based control guarantees the stability of closed loop systems. However, applying the DO design to the LMI framework is not trivial because there are two control targets, whereas the standard LMI stabilizes a single control target. In this study, the problem is first resolved by building a single fictitious model because the two models are serial and can be considered as a single model from the Q-filter point of view. Using the proposed design framework, all-stabilizing Q filters are calculated. In addition, for the stability and robustness of the DO, two metrics are proposed to quantify the stability and robustness and combined into a single unified index to satisfy both metrics. Based on an application example, it is verified that the proposed method is effective, with a performance improvement of 10.8%.


2005 ◽  
Vol 04 (01) ◽  
pp. 117-126
Author(s):  
N. L. MA ◽  
P. WU

Using density functional theory, we predicted the solution structure of the hydrolyzed 3–aminopropyltriethoxysilane (h–APS), which is a silane coupling agent commonly used in many industrial applications. We have located five stable minima on the potential energy surface of h–APS in which four of them are "neutral", and the remaining one is zwitterionic (dipolar) in nature. Our calculations suggested that the stability of the most stable form of h–APS in water (denoted as II_N) arose from strong intramolecular OH ⋯ N hydrogen bond. The least stable form is the zwitterionic form (I_ZW), which is estimated to be over 90 kJ mol -1 less stable than II_N. The factors governing the relative stabilities of different forms are discussed.


2013 ◽  
Vol 6 (11) ◽  
pp. 876-883 ◽  
Author(s):  
MBS Donio ◽  
SFA Ronica ◽  
V Thanga Viji ◽  
S Velmurugan ◽  
J Adlin Jenifer ◽  
...  

2018 ◽  
Vol 41 (8) ◽  
pp. 2352-2364 ◽  
Author(s):  
Arif Iqbal ◽  
Girish Kumar Singh

Owing to the superior properties and stable operation, the Permanent Magnet Synchronous Motor (PMSM) is preferably used in wide industrial applications. But, the stability of motor is found to be dependent on its initial operating condition, showing the chaotic characteristic. Therefore, this paper addresses the chaos control of PMSM by developing four simple but effective controllers, which are mathematically designed by using the principle of Lyapunov’s method for asymptotic global stability. A comparative performance assessment has been carried out for the developed controllers in terms of settling time and peak over shoot. Furthermore, the concept of conventional proportional-integration type controller has been extended to develop two more controllers for chaos control of PMSM. Numerical simulation has been carried out in Matlab environment for performance evaluation of developed controllers. The obtained analytical results have been validated through experimental implementation in real time environment on Multisim/Ultiboard platform.


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