Isolation and characterization of alkaline protease bacteria from sheep leather.

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Journal name: World Journal of Pharmaceutical Research
Original article title: Isolation, optimization and characterization of alkaline protease producing bacteria from collagen layer of decaying skin (sheep upper ) leather industry
The WJPR includes peer-reviewed publications such as scientific research papers, reports, review articles, company news, thesis reports and case studies in areas of Biology, Pharmaceutical industries and Chemical technology while incorporating ancient fields of knowledge such combining Ayurveda with scientific data.
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Author:

S.Manikandan, Dr.P.Kavitha, Dr.A.Panneerselvam


World Journal of Pharmaceutical Research:

(An ISO 9001:2015 Certified International Journal)

Full text available for: Isolation, optimization and characterization of alkaline protease producing bacteria from collagen layer of decaying skin (sheep upper ) leather industry

Source type: An International Peer Reviewed Journal for Pharmaceutical and Medical and Scientific Research


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Summary of article contents:

Introduction

Enzymes are essential biological catalysts that facilitate numerous chemical reactions vital for life, playing significant roles in various industries such as pharmaceuticals, food, and detergents. Proteases, a class of enzymes capable of breaking down proteins into peptides and amino acids, are particularly relevant in biotechnological processes. Among proteases, alkaline proteases are significant due to their stability in alkaline environments, making them useful in various applications, including detergent formulations and leather processing. This study focuses on isolating and characterizing alkaline protease-producing bacteria from decaying sheep upper leather, emphasizing the implications of these findings for industrial applications.

Alkaline Proteases in Industry

Alkaline proteases, also known as subtilisins, are commercially essential enzymes primarily used for their ability to hydrolyze proteins in alkaline conditions. The global market for industrial enzymes has grown significantly, with alkaline proteases accounting for a substantial portion of this market. Their use in detergents, for instance, allows for effective stain removal at lower temperatures, making laundry processes more energy-efficient. The study highlights the potential economic benefits of utilizing alkaline proteases, particularly as industries strive for sustainability and reduced chemical dependency.

Isolation and Screening of Bacteria

In the research, 21 bacterial strains were isolated from the collagen layer of decaying sheep leather, with 15 exhibiting proteolytic activity on skim milk agar medium at alkaline pH. The identified genera of bacteria were primarily Bacillus and Pseudomonas, known for their ability to produce alkaline proteases. The study employed selective media and biochemical tests to characterize the strains, indicating that samples collected from various tanneries yielded notable enzyme activity. This highlights the potential of utilizing waste materials from the leather industry as a source for microbial enzyme production.

Optimization of Enzyme Production

The researchers optimized several parameters influencing alkaline protease production, including pH, temperature, incubation time, and inoculum size. The highest enzyme yield was achieved at pH 10 and a temperature of 37°C, indicating that environmental conditions significantly impact microbial enzyme performance. Time optimization studies suggested that protease levels increased with longer incubation periods, although a decline was observed after 48 to 60 hours, suggesting potential autolysis or resource depletion. Additionally, the optimal inoculum size for effective enzyme production was determined to be 1 ml, balancing sufficient microbial presence for enzyme secretion without over-crowding.

Conclusion

The findings of this study underscore the importance of alkaline protease-producing bacteria from leather processing waste, particularly those belonging to the Bacillus genus. The successful isolation and characterization of these microbial strains demonstrate their potential to replace harmful chemicals currently used in various industries, leading to more sustainable practices. Given the increasing demand for environmentally friendly alternatives, further research into optimizing growth conditions and exploring commercial applications for these enzymes could significantly impact the detergent and leather industries. Overall, this investigation contributes to the ongoing efforts to harness microbial biodiversity for industrial use, presenting a pathway toward sustainable enzyme production.

FAQ section (important questions/answers):

What is the significance of alkaline protease in industries?

Alkaline protease is essential in industries like detergents, food, pharmaceuticals, and leather. It helps in protein degradation, making processes more efficient and reducing reliance on harmful chemicals.

How were bacterial strains isolated for this study?

Bacterial strains were isolated from decaying sheep leather samples collected from various tanneries. Dilutions of skin samples were inoculated on nutrient agar plates adjusted to pH 9.

Which microorganisms were identified in the study?

The study identified several bacterial strains, primarily belonging to the genera Bacillus and Pseudomonas, known for producing alkaline proteases.

What factors influenced alkaline protease production in bacteria?

Production was influenced by pH, incubation time, temperature, and inoculum size. Optimal conditions varied, with the best results found at pH 10 and 37°C.

What method was used to measure proteolytic activity?

Proteolytic activity was measured using a casein solution and monitoring the release of tyrosine at specific conditions, determining enzyme activity based on absorbance.

What are the potential environmental benefits of using proteases?

Using proteases in industrial applications can replace harmful chemicals, decreasing environmental pollution and promoting safer, more sustainable production methods.

Glossary definitions and references:

Scientific and Ayurvedic Glossary list for “Isolation and characterization of alkaline protease bacteria from sheep leather.”. This list explains important keywords that occur in this article and links it to the glossary for a better understanding of that concept in the context of Ayurveda and other topics.

1) Medium:
In the context of microbiology, 'medium' refers to the nutrient-rich environment where bacteria or other microorganisms grow. The choice of medium, including its pH and composition, is critical for promoting specific microbial activity, such as protease production. In the study, skim milk agar was utilized to observe proteolytic activity.

2) Activity:
The term 'activity' signifies the functional capacity of enzymes, in this case, proteases, to catalyze reactions. It reflects the enzyme's efficiency in breaking down proteins into peptides and amino acids. Monitoring enzymatic activity through experiments is vital for quantifying the performance of isolated bacterial strains.

3) Table:
In research documentation, a 'table' presents organized data, such as experimental results or characteristics of isolated bacterial strains. Tables facilitate comparison and analysis by summarizing complex information succinctly, making it easier for readers to extract essential findings from the study’s results on protease-producing bacteria.

4) Study (Studying):
The word 'study' indicates a systematic investigation aimed at gaining new knowledge or confirming previously known facts. In this context, the study focused on isolating, optimizing, and characterizing alkaline protease-producing bacteria from the collagen layer of decaying sheep skin, showcasing its relevance to industrial applications.

5) Soil:
Soil serves as an ecological reservoir for diverse microbial communities, including alkalophiles, which thrive in specific pH conditions. In the study, normal garden soil was referenced as a habitat for isolating protease-producing bacteria, emphasizing the significance of soil environments in microbial biodiversity and enzyme production.

6) Tamilnadu (Tamil-nadu, Tamilnāḍu):
'Tamil Nadu' is a state in southern India and is central to this study as the geographical location from where samples were collected. The environmental conditions in Tamil Nadu, particularly in relation to leather processing and biodiversity, influence the microbial communities and their enzymatic activities described in the research.

7) Chennai:
Chennai, the capital city of Tamil Nadu, is mentioned as one of the locations where the study collected sheep skin samples from tanneries. The urban environment of Chennai represents an intersection of natural and industrial influences on microbial diversity, particularly concerning the productivity of enzymes in tannery waste.

8) Company:
'Company' refers to commercial entities engaged in leather processing. In this context, various tannery companies in Tamil Nadu, such as Naser Tanning Company, provided the collagen sample for isolating protease-producing bacteria, illustrating the relationship between industry and microbial research in enzyme production.

9) Tamil (Tamiḻ):
The term 'Tamil' pertains to the Dravidian ethnicity and language spoken predominantly in Tamil Nadu. It signifies the cultural context and region of the study, framing the local practices in the leather industry and the biodiversity explored in relation to alkaline protease-producing microorganisms.

10) Nadu (Nāḍu):
'Nadu', meaning 'land' in Tamil, is part of the name 'Tamil Nadu'. It emphasizes the regional focus of the research, grounding the ecological and industrial conditions relevant to the study of microbial enzymatic activity. Understanding local environmental factors is crucial for optimizing enzyme production.

11) Bhavani (Bhavāni, Bhavānī):
Bhavani is a town in Tamil Nadu where one of the tanneries contributing to the study's raw material was located. Its mention highlights the local industrial practices in the leather industry, essential for understanding the microbial diversity and enzymatic production studied in the research.

12) Trichy:
Trichy, another significant city in Tamil Nadu, is noted for its tannery operations. The inclusion of Trichy situates the research within the leather processing context of Tamil Nadu, emphasizing the geographical diversity and the importance of multiple sampling sites for isolating protease-producing bacteria.

13) Milk:
In microbiology, 'milk' often denotes a nutrient source used in culture media for growing bacteria, particularly in assays to test for proteolytic activity. The use of skim milk agar in this study highlights its role as a substrate to identify protease-producing microorganisms effectively.

14) Food:
The term 'food' refers to organic substances consumed by living organisms for nutrition. In this study, proteases derived from microbial sources may have implications in food processing, particularly in breaking down proteins to enhance the digestibility and flavor of food products, showcasing their versatility.

15) Species:
'Species' is a fundamental taxonomic category that groups organisms sharing common characteristics. In this research, specific microbial species, particularly strains of Bacillus and Pseudomonas, were identified as key producers of alkaline proteases, crucial for applications in industries like leather and detergents.

16) Life:
The term 'life' encompasses all living organisms, including microorganisms responsible for vital biochemical processes. The study underscores the relationship between microbial life, particularly protease-producing bacteria, and industrial applications, revealing the potential for biotechnological exploitation of these organisms.

17) Thanjavur:
Thanjavur is a city in Tamil Nadu where the study's research institution is located. Its mention contextualizes the geographical and cultural background of the research, contributing to the understanding of local biodiversity, specifically within the framework of the leather industry and enzyme production.

18) Habitat:
In ecological terms, 'habitat' refers to the environment where organisms live. The habitats of alkalophilic microorganisms discussed in the study include man-made alkaline locations such as tanneries and natural environments, which are critical for understanding their adaptability and the associated enzymatic activities.

19) Botany:
Botany, the scientific study of plants, is the broader field within which this research is situated. Although primarily focused on microbiology, the study's affiliation with the Department of Botany signifies an interdisciplinary approach, bridging botanical sciences and microbial enzyme research for industrial applications.

20) Smita:
Smita refers to one of the authors involved in the research study. The acknowledgment of contributors highlights the collaborative effort in the scientific community, emphasizing that the findings are a result of collective expertise and research experience in the areas of microbiology and enzyme production.

21) India:
'India' denotes the country where the research was conducted, framing the study within its national ecological and industrial context. Understanding the local biodiversity and industrial practices in India is crucial for evaluating the potential for microbial exploitation in enzyme production for various industries.

22) Water:
'Water' is a critical component in microbial processes, serving as a medium in which biochemical reactions occur. In this study, it is indirectly referenced through the preparation of culture media and the importance of maintaining optimal water conditions for microbial growth and enzyme activity.

23) Observation:
The term 'observation' pertains to the systematic monitoring of experimental phenomena. In this study, observations made during enzymatic assays and bacterial screenings were essential for characterizing the microbial isolates and determining their proteolytic activities, ultimately guiding the research findings and conclusions.

24) Discussion:
In academic research, 'discussion' refers to the section where findings are interpreted in the context of existing knowledge. This portion of the study critically analyzes the results obtained from experiments, positions them within the broader scientific framework, and explores their implications for future research and practical applications.

25) Gelatin:
Gelatin is a protein derived from collagen, often used in microbiological media as a solidifying agent. Its mention in this study signifies the relevance of protein substrates in evaluating proteolytic activities, reflecting the importance of such compounds in testing the capabilities of protease-producing bacteria.

26) Forest:
The term 'forest' implies a natural environment where diverse plant and animal life thrives. Although not directly studied here, forests may harbor unique microbial biodiversity, suggesting potential for discovering new enzymes, including proteases, that could be exploited for industrial purposes, paralleling the exploration conducted in this research.

27) Nature:
'Nature' encompasses all living organisms and the ecosystems they inhabit. In the study, the exploration of microbial life, particularly the enzymes they produce, reflects a broader understanding of the natural world, emphasizing the significance of biotechnological applications derived from biological processes inherent in nature.

28) Reason:
The term 'reason' signifies the rationale behind specific choices made in research, such as opting for alkaline protease production from certain bacterial strains. Understanding the reasons for selecting particular microbes or methodologies plays a crucial role in optimizing experimental outcomes and achieving practical applications.

29) Animal:
The term 'animal' refers to multicellular organisms that can contribute to a variety of ecological roles, including as sources for microbial isolation. In this study, sheep skin was specifically utilized, illustrating the intersection between animal by-products and the search for identifying protease-producing microorganisms.

30) Egypt:
'Egypt' is referenced in relation to specific ecological contexts that harbor unique microbial populations. In the study, it suggests that extreme environments, such as those found in Egypt, could potentially yield unique isolates that might share similarities with organisms studied from the decay of leather in Tamil Nadu.

31) Joshi (Jōśī, Jośī):
Joshi, likely a reference to one of the authors or researchers, signifies the collaborative nature of scientific inquiry. It underscores the importance of multiple contributions and insights from various experts, which are essential for enriching the findings of studies like the one focusing on protease-producing bacteria.

32) Kumar (Kumār):
Kumar typically represents a key contributor or researcher within the study, indicating collaborative research efforts essential for scientific advancement. Through collaboration, research efforts can lead to more comprehensive findings, particularly concerning the relationship between microbial activity and industrial applications.

33) Sudha (Sudhā):
Sudha is referenced as one of the co-authors of the study, highlighting the multi-author collaboration needed in scientific research. The inclusion of different perspectives and expertise in studies assists in achieving a thorough examination of the results obtained in studying protease-producing bacteria.

34) Jaya (Jāyā, Jayā):
Jaya signifies an individual possibly involved in the methodology or research aspect of the study as an author. Their contributions help shape the reliability and depth of the research findings, underlining the collaborative effort integral to advancing knowledge in microbiology and enzyme production.

35) Hand:
In this context, 'hand' could refer to the human involvement in scientific observation and experimentation, emphasizing the role of researchers in meticulously conducting the study. Human intervention is critical in isolating and characterizing microbial strains, determining their activities, and applying findings in real-world scenarios.

36) Drug:
The term 'drug' refers to substances used for medical purposes, where enzymes from microorganisms could play a crucial role in drug production or processing. This highlights the potential applicability of microbial proteases in pharmaceuticals, illustrating the broader implications of enzyme research in medicinal development.

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