ABSTRACT
To date, there have been no or just a few reports of successful cloning and expression to create biologically active ocins or bacteriocins. Cloning, expression, and production of class I ocins are problematic because of their structural arrangements, coordinated functions, size, and posttranslational modifications. Mass synthesis of these molecules is necessary for commercialization and to restrict the excessive use of conventional antibiotics, which encourages the development of antibiotic-resistant bacteria. In the case of class III ocins, there are no reports of obtaining biological active proteins to date. Being able to obtain biologically active proteins requires an understanding of mechanistic features due to their expanding importance and broad spectrum of activity. As a result, we intend to clone and express the class III type. The class I types that are devoid of posttranslational modifications were transformed into class III through fusion. Therefore, this construct resembles a class III type ocin. With the exception of Zoocin, expression of the proteins was found to be physiologically ineffective after cloning. But, few cell morphological changes such as elongation, aggregation, and the formation of terminal hyphae were observed. However, it was discovered that the target indicator had been altered to Vibrio spp. in a few. All the three ocins were subjected to in-silico structure prediction/analysis. Finally, we confirm the existence of unidentified additional intrinsic factors for successful expression to obtain biologically active protein.
Subject(s)
Bacteriocins , Amino Acid Sequence , Bacteriocins/genetics , Bacteriocins/chemistry , Anti-Bacterial Agents/chemistry , Transcription FactorsABSTRACT
An antimicrobial peptide [Bacillus antimicrobial peptide (BAMP)] produced by Bacillus paralicheniformis was isolated from the Indian traditional fermented food and characterized. The antimicrobial peptide BAMP showed many unique features such as thermostability (4.0-125°C), pH tolerance (pH 2.0-9.0), and resistance to physiological enzymes (trypsin, chymotrypsin, pepsin, proteinase K, protease, and catalase), and food-grade metal salts do not inhibit the activity. The broad spectrum of BAMP (antimicrobial activity) makes it a suitable candidate for food preservation as well as antimicrobial therapy. BAMP was found to exhibit a bacteriostatic effect on Salmonella typhi and controls the viability of Listeria monocytogenes in chicken meat efficiently. BAMP was found to establish eubiosis, as it is not antagonistic to Lactobacillus. Its non-hemolytic nature makes it suitable for therapy. Various genome prediction tools were adopted and applied to understand their localization, gene arrangement, and type of antimicrobials. Founded on its superior functional attributes, BAMP is a potent new-generation antimicrobial peptide.
ABSTRACT
The food industry produces highly perishable products. Food spoilage represents a severe problem for food manufacturers. Therefore, it is important to identify effective preservation solutions to prevent food spoilage. Ocins (e.g., bacteriocins, lactocins, and enterocins) are antibacterial proteins synthesized by bacteria that destroy or suppress the growth of related or unrelated bacterial strains. Ocins represent a promising strategy for food preservation, because of their antagonist effects toward food spoilage microorganisms, high potency, and low toxicity. Additionally, they can be bioengineered. The most common and commercially available ocins are nisin, plantaracin, sakacin P, and pediocin. Several ocins have been characterized and studied biochemically and genetically; however, their structure-function relationship, biosynthesis, and mechanism of action are not understood. This narrative review focuses primarily on ocins and their relevance to the food industry to help prevent food spoilage. In particular, the applications and limitations of ocins in the food industry are highlighted.
ABSTRACT
The ocins are antimicrobial polypeptides produced by probiotic microbes, such as Lactobacillus , Enterococcus , Streptococcus , Leuconostoc and Bifidobacterium . They are produced in response to stress and for the self-defense of the bacterium. It is indispensable to understand their mechanistic characteristics, structures, and functions, if the food industry is to reduce contamination levels and produce germfree foods. Databases of the ocins that are readily accessible to the food industry are scarce, but urgently required. Therefore, we established a very useful, unique, and a simple ocin database, which not merely provides information about ocins, but also directs their utilisation in the food industry. The database includes information about each ocin, its amino acid sequence, molecular weight, and isoelectric point. The database also possesses all the currently known ocin (probiotic origin only) sequences and structures, target organisms, and relevant to food industries (aqua culture, dairy and meat industries), which is hard to obtain in other databases. The database is free for public and accessed at http://ocins.cftri.com/ocins/.
ABSTRACT
The generation of a zone of inhibition on a solid substrate indicates the bioactivity of antimicrobial peptides such as bacteriocin and enterocin. The indicator strain plays a significant role in bacteriocin assays. Other characteristics of bacteriocins, such as their dispersal ability and the different zymogram components, also affect bacteriocin assays. However, universal well diffusion assays for antimicrobials, irrespective of their ability to diffuse (bacteriocin and enterocin), do not exist. The ability of different zymography components to generate non-specific activities have rarely been explored in the literature. The purpose of the present work was to evaluate the impact of major factors (diffusion and rate of diffusion) in a solid substrate bioassay, and to document the adverse effects of sodium dodecyl sulfate in zymograms used to estimate the approximate molecular weight of bacteriocins.
