scholarly journals Isolation, Screening, Characterization and Identification of Alkaline Protease Producing Bacteria from Leather Industry Effluent

2021 ◽  
Author(s):  
Chandran Masi ◽  
Getachew Gemechu ◽  
Mesfin Tafesse
Keyword(s):  
Alkaline Protease ◽  
Protease Activity ◽  
Bacterial Species ◽  
Skim Milk ◽  
Industrial Effluents ◽  
Leather Industry ◽  
Skim Milk Agar ◽  
Secondary Screening ◽  
Agar Media ◽  
Industry Effluent

Abstract BackgroundA wide variety of Bacterial species produces protease enzyme and the application of same enzyme have been manipulated precisely and used in various biotechnological areas including industrial and environmental sectors. The main aim of this research study was to isolate, screen and identify protease producing bacteria which were sampled from leather industry effluent present in the outer skirts of Addis Ababa, Ethiopia.PurposeTo isolated alkaline protease producing bacteria from leather industrial effluents and to characterization (Secreening and identification).MethodsSample collected from Modji leather industrial effluents and stored in the microbiology lab. After isolated bacteria from effluent using serial dilution and followed by isolate protease producing bacteria using skim milk agar media. After studying Primary and secondary screening using zonal inhibition methods to select potential protease producing bacteria using skim milk agar media. Finally to characterization and identification of potential bacteria using biochemical methods, protein estimation, biomass, protease assay and gene sequencing (16S rRNA) method to finalized best protease producing bacteria. ResultsTwenty-eight different bacterial colonies were isolated initially from the leather industry effluent sample situated at Modjo town of Ethiopia. The isolated bacteria were screened using primary screening method with skim milk agar medium. Three isolates namely MS12, ML5 and ML12 showing highest zone of proteolysis as a result of casein degradation on the agar plates were selected and subjected to secondary screening. Further secondary secreening confirmed that MS12, ML5 and ML12 has efficient proteolytic activity and can be considered as potent protease producer. The three isolates were then subjected to morphological and biochemical tests to identify probably bacterial species and all the three bacterial isolates were found out to be of Bacillus species. Shake flask method was carried out to identify the most potent one having greater biomass production capabilities, protein quantity and protease activity. ML12 isolated from leather effluent waste showed highest Protein(170mg/ml), Protease activity(19U/ml), high biomass production and the same was subjected to molecular identification using 16s sequencing and a Phylogenetic tree was constructed to identify the closest neighbor. The isolate ML12 is 97.87% homologous to Bacillus cereus strain (KY995152.1) and 97.86% homologous to Bacillus cereus strain (MK968813.1).ConclusionsThis study has revealed that the leather industry effluent site has significant feature of housing potent bacterial species producing protease of commercial value. Being one among the most widely used enzyme, comparatively. Protease holds a larger scope for research and commercialization any other type of enzymes. There is a need to develop novel protease enzymes for further necessary applications of these enzymes. Moreover, enzyme produced by bacteria which are present in effluents are a greater boon to establish the significance of converting industrial wastes to a highly valuable enzymes especially like proteases.

scholarly journals Isolation, screening, characterization, and identification of alkaline protease-producing bacteria from leather industry effluent

2021 ◽  
Vol 71 (1) ◽  
Author(s):  
Chandran Masi ◽  
Getachew Gemechu ◽  
Mesfin Tafesse
Keyword(s):  
Bacillus Cereus ◽  
Alkaline Protease ◽  
Protease Activity ◽  
Bacterial Species ◽  
Skim Milk ◽  
Industrial Effluents ◽  
Leather Industry ◽  
Skim Milk Agar ◽  
Secondary Screening ◽  
Industry Effluent

Abstract Background A wide variety of bacterial species produces protease enzyme, and the application of the same enzyme has been manipulated precisely and used in various biotechnological areas including industrial and environmental sectors. The main aim of this research study was to isolate, screen, and identify alkaline protease-producing bacteria that were sampled from leather industry effluent present in the outer skirts of Addis Ababa, Ethiopia. Purpose To isolate and characterize the alkaline protease-producing bacteria from leather industrial effluents. Methods Samples are collected from Modji leather industrial effluents and stored in the microbiology lab. After isolated bacteria from effluent using serial dilution and followed by isolated protease-producing bacteria using skim milk agar media. After studying primary and secondary screening using zonal inhibition methods to select potential protease-producing bacteria using skim milk agar media. Finally, to identify the potential bacteria using biochemical methods, bacterial biomass, protease activity, and gene sequencing (16S rRNA) method to finalize the best alkaline protease producing bacteria identified. Results First twenty-eight different bacterial colonies were isolated initially from the leather industry effluent sample situated at the Modjo town of Ethiopia. The isolated bacteria were screened using the primary and secondary screening method with skim milk agar medium. At the primary level, we selected three isolates namely ML5(14 mm), ML12(18 mm), and MS12 (15 mm), showing the highest zone of proteolysis as a result of casein degradation on the agar plates were selected and subjected to primary screening. Further secondary screening confirmed that the zone of inhibition methods ML5 (14.00±0.75 mm), ML12 (19.50±0.66 mm), and MS12 (15.00±1.32 mm) has efficient proteolytic activity and can be considered as effective protease producer. The three isolates were then subjected to morphological and biochemical tests to identify probably bacterial species, and all the three bacterial isolates were found out to be of Bacillus species. The shake flask method was carried out to identify the most potent one having greater biomass production capabilities and protease activity. ML12 isolated from leather effluent waste showed the highest protease activity (19 U/ml), high biomass production, and the same was subjected to molecular identification using 16s sequencing and a phylogenetic tree was constructed to identify the closest neighbor. The isolate ML12 (Bacillus cereus strain -MN629232.1) is 97.87% homologous to Bacillus cereus strain (KY995152.1) and 97.86% homologous to Bacillus cereus strain (MK968813.1). Conclusions This study has exposed that from twenty-eight different bacterial samples isolated from leather industry effluent; further primary and secondary screening methods were selected three potential alkaline protease strains. Finally, based on its biochemical identification, biomass, and protease activity, ML12 (Bacillus cereus strains) is the best strain identified. The alkaline protease has the significant feature of housing potent bacterial species for producing protease of commercial value.

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%.

scholarly journals Comparative Effects of Industrial Effluents and sub-metropolitan Sewage of Biratnagar on Germination and seedling growth of Rice and Blackgram

Our Nature ◽  
1970 ◽  
Vol 1 (1) ◽  
pp. 10-14 ◽  
Author(s):  
Bhabindra Niroula
Keyword(s):  
Toxic Effect ◽  
Seedling Growth ◽  
Industrial Effluents ◽  
Lethal Effect ◽  
Toxic Effects ◽  
Black Gram ◽  
Leather Industry ◽  
Iron And Steel ◽  
Power House ◽  
Industry Effluent

Comparative effects of effluents from six major industries viz. Diesel Power House, Hetaunda Iron and Steel, Hulas Wire, Himalaya Soap and Chemicals, Leather Industry, Shah Udyog and sub-metropolitan Sewage of Biratnagar on germination and seedling growth of rice and black gram were studied. Effluent of Himalaya Soap and Chemicals showed toxic lethal effect on both the test crops. On germination rice remained more sensitive and susceptible to the toxic effects of industrial effluents but black gram proved to be more tolerant. Effluents of Diesel power House and Shah Udyog remained toxic for seedling growth of black gram as their effects were significant while Leather Industry effluent showed toxic effect on rice for germination as well as seedling growth. Keywords: Industrial effluents, Sewage, Germinationdoi:10.3126/on.v1i1.296Our Nature (2003) 1: 10-14

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.

Resistance of Phlebotomus papatasi to infection with Leishmania donovani is modulated by components of the infective bloodmeal

Parasitology ◽  
1998 ◽  
Vol 117 (5) ◽  
pp. 467-473 ◽  
Author(s):  
Y. SCHLEIN ◽  
R. L. JACOBSON
Keyword(s):  
Proteolytic Activity ◽  
Trypsin Inhibitor ◽  
Alkaline Protease ◽  
Protease Activity ◽  
Leishmania Donovani ◽  
Blood Feeding ◽  
Growth Conditions ◽  
Enzymatic Activities ◽  
Phlebotomus Papatasi ◽  
Proteolytic Activities

The circumstances which permit the establishment of Leishmania infections in sandflies were investigated by altering the growth conditions for L. donovani parasites in the unsuitable vector Phlebotomus papatasi. Only 5·0% of the sandflies harboured a few parasites 3 days after feeding on promastigotes in defibrinated blood. Heparinized blood or the addition of trypsin inhibitor to the meals allowed persistence of infections (day 6) in 9·9% and 25·8% of the flies respectively. Meals of erythrocytes, saline and amastigotes produced 44·4% fly infection on day 6, while similar promastigote-initiated infections remained in 70·3% of the flies. Proteolytic activities in the guts of sandflies fed on the above meals without parasites, were the highest after defibrinated bloodmeals. Erythrocytes with saline decreased the maximal alkaline protease level from 20·8 U to 13·5 U/fly; that of trypsin from 3·9 U to 1·8 U/fly and that of the aminopeptidase from 5·5 U to 3·9 U/fly. After meals of heparinized blood, the maximal alkaline protease activity (12·0 U/fly) was also much lower than after defibrinated blood-feeding. The different diets which resulted in comparatively low enzymatic activities, including blood with trypsin inhibitor, also promoted the survival of infections. This implies that the proteolytic activity in the sandfly gut modulates the vector susceptibility.

Sewage water irrigation effects on some potentially toxic trace elements in soil and potato plants in northwestern India

2000 ◽  
Vol 80 (3) ◽  
pp. 465-471 ◽  
Author(s):  
M. S. Brar ◽  
S. S. Malhi ◽  
A. P. Singh ◽  
C. L. Arora ◽  
K. S. Gill
Keyword(s):  
Solanum Tuberosum ◽  
Ground Water ◽  
Toxic Elements ◽  
Crop Yields ◽  
Solanum Tuberosum L ◽  
Sewage Water ◽  
Potentially Toxic Elements ◽  
Leather Industry ◽  
Northwestern India ◽  
Industry Effluent

Sewage water is used for irrigation to improve crop yields, but it may affect the soil and crop quality. The effects of discharging the effluents of a leather complex on the concentrations of some potentially toxic elements (Cu, Fe, Mn, Zn, Al, As, Cr and Ni) in sewage water and the effects of irrigation with contaminated sewage water on the concentrations of these elements in soils and in potato (Solanum tuberosum L.) leaves and tubers were studied near Jalandhar city, Punjab, in northwestern India. Two treatments were 10 yr of irrigation with 300 mm of sewage water + 300 mm of ground water per annum (SW) and with 600 mm of ground water per annum (GW). Soils of the study fields were Typic Ustochripts with sandy to loamy sand texture. The concentrations of all elements except As increased following the addition of leather complex effluents in the sewage water, with the biggest increase in the concentration of Cr (from 2.7 mg to 14.0 mg Cr L−1). The SW treatment increased concentrations of all elements in soil except As, and the increase was significant to 60 cm depth for Fe, Mn, Zn, Al and Ni and to 30 cm depth for Cu and Cr. Irrigation with SW also increased the concentrations of these elements in potato leaves and tubers and the increase was generally higher in leaves than in tubers. The proportional increase of Cu, Fe, Zn and Al was less in plants than in soils, that of Mn and Cr was almost similar in plants and in soil, and that of Ni was more in plants than in soil. These elements, which accumulate in soils and crops, may become health hazards to humans and/or animals. Therefore, continued monitoring of the concentrations of potentially toxic elements in soil and plants and/or treatment of sewage water before using for irrigation is needed. Key words: Irrigation, leather industry effluent, Solanum tuberosum, potentially toxic elements, sewage water

scholarly journals Comparison between freeze and spray drying to obtain powder Rubrivivax gelatinosus biomass

2013 ◽  
Vol 33 (1) ◽  
pp. 47-51 ◽  
Author(s):  
Edson Francisco do Espírito Santo ◽  
Leandro Kanamaru Franco de Lima ◽  
Ane Pamela Capucci Torres ◽  
Gabriela de Oliveira ◽  
Elisa Helena Giglio Ponsano
Keyword(s):  
Spray Drying ◽  
Freeze Drying ◽  
Bacterial Biomass ◽  
Quality Criterion ◽  
Industrial Effluents ◽  
Product Recovery ◽  
Animal Origin ◽  
Freeze Dried ◽  
Rubrivivax Gelatinosus ◽  
Industry Effluent

The use of colorants in products of animal origin is justified by the improvement in the color of foods since this attribute is considered a quality criterion. These additives can be produced using industrial effluents as substrates and appropriate organisms, such as Rubrivivax gelatinosus. Oxycarotenoids represent a class of carotenes responsible for the pigmentation of animals and vegetables. R. gelatinosus grows in fish industry effluent with the resulting production of a bacterial biomass containing oxycarotenoids. The purpose of this study was to compare the use of two drying processes - spray and freeze drying - to obtain powder biomass in terms of the process parameters (yield, productivity, and product recovery) and the product characteristics (color, proximate composition, and oxycarotenoids). No difference was detected in the yield between these techniques, while productivity was higher using spray drying. Higher product recovery and moisture were achieved with freeze drying, while ash was higher with spray drying. The freeze dried biomass was redder, darker and less saturated than the spray dried biomass. No difference in oxycarotenoids was detected between the biomasses. Although it results in lower recovery rate, spray drying was faster and more productive, and it provided the same yield as freeze drying, which makes it the method of choice for obtaining R. gelatinosus biomass.

scholarly journals Production of fibrinogenolytic and fibrinolytic enzymes by a strain of Penicillium sp. isolated from contaminated soil with industrial effluent

2019 ◽  
Vol 41 (1) ◽  
pp. 40606 ◽  
Author(s):  
Lillian Maria Baggio ◽  
Luciano Aparecido Panagio ◽  
Fabiana Guillen Moreira Gasparin ◽  
Daniele Sartori ◽  
Maria Antonia Pedrine Colabone Celligoi ◽  
...  
Keyword(s):  
Cardiovascular Diseases ◽  
Industrial Effluent ◽  
Skim Milk ◽  
Industrial Effluents ◽  
Media Composition ◽  
Lytic Enzymes ◽  
Fibrinolytic Enzymes ◽  
Health Area ◽  
Tissue Plasminogen ◽  
Selection Of

Cardiovascular diseases associated with thrombosis are one of the main causes of death all around the world. Urokinase, streptokinase, and tissue plasminogen activator are the major thrombolytic agents used to treat thrombosis. However, the fact that these agents have several side effects and high prices has driven the search for safer and more economically viable compounds for the treatment of cardiovascular diseases. Thus, the aim of this study was to evaluate the potential of fungi isolated from industrial effluents to produce fibrino(geno)lytic enzymes. The selection of the protease-producing strains showed that only the BF20 strain was able to produce proteolytic halos in milk agar. This strain identified as belonging to the genus Penicillium was cultivated in submerged fermentation. Different media composition were tested to evaluate proteolytic activity, and the results showed that the medium containing 0.1% yeast extract and 1% skim milk, pH 5.0, present higher azocaseinolytic activity (0.24 U mL-1 min.-1). This sample also showed the ability to degrade fibrinogen and fibrin after 15 and 120 min. of incubation, respectively. These results indicate that the BF20 strain has a thrombolytic potential, effectively degrading fibrinogen and fibrin, having great application in the health area.

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