Production, Purification, and Biochemical Characterization of Thermostable Metallo-Protease from Novel Bacillus alkalitelluris TWI3 Isolated from Tannery Waste.

Protease enzymes in tannery industries have enormous applications. Seeking a potential candidate for efficient protease production has emerged in recent years. In our study, we sought to isolate proteolytic bacteria from tannery waste dumping site in Tamilnadu, India. Novel proteolytic Bacillus alkalitelluris TWI3 was isolated and tested for protease production. Maximum protease production was achieved using lactose and skim milk as a carbon and nitrogen source, respectively, and optimum growth temperature was found to be 40 °C at pH 8. Protease enzyme was purified using ammonium sulfate precipitation method and anion exchange chromatography. Diethylaminoethanol (DEAE) column chromatography and Sephadex G-100 chromatography yielded an overall 4.92-fold and 7.19-fold purification, respectively. The 42.6-kDa TWI3 protease was characterized as alkaline metallo-protease and stable up to 60 °C and pH 10. Ca(2+), Mn(2+), and Mg(2+) ions activated the protease, while Hg(2+), Cu(2+), Zn(2+), and Fe(2+) greatly inhibited it. Ethylenediaminetetraacetic acid (EDTA) inhibited TWI3 protease and was activated by Ca(2+), which confirmed that TWI3 protease is a metallo-protease. Moreover, this protease is capable of dehairing goat skin and also removed several cloth stains, which makes it more suitable for various biotechnological applications.

Determining the probiotic potential of cholesterol-reducing Lactobacillus and Weissella strains isolated from gherkins (fermented cucumber) and south Indian fermented koozh.

This study sought to evaluate the probiotic potential of lactic acid bacteria (LAB) isolated from traditionally fermented south Indian koozh and gherkin (cucumber). A total of 51 LAB strains were isolated, among which four were identified as Lactobacillus spp. and three as Weissella spp. The strains were screened for their probiotic potential. All isolated Lactobacillus and Weissella strains were capable of surviving under low pH and bile salt conditions. GI9 and FKI21 were able to survive at pH 2.0 and 0.50% bile salt for 3 h without losing their viability. All LAB strains exhibited inhibitory activity against tested pathogens and were able to deconjugate bile salt. Higher deconjugation was observed in the presence of sodium glycocholate (P < 0.05). Strain FKI21 showed maximum auto-aggregation (79%) and co-aggregation with Escherichia coli MTCC 1089 (68%). Exopolysaccharide production of LAB strains ranged from 68.39 to 127.12 mg/L (P < 0.05). Moreover, GI9 (58.08 µg/ml) and FKI21 (56.25 µg/ml) exhibited maximum cholesterol reduction with bile salts. 16S rRNA sequencing confirmed GI9 and FKI21 as Lactobacillus crispatus and Weissella koreensis, respectively. This is the first study to report isolation of W. koreensis FKI21 from fermented koozh and demonstrates its cholesterol-reducing potential.

An ethnobotanical study of indigenous knowledge on medicinal plants used by the village peoples of Thoppampatti, Dindigul district, Tamilnadu, India.

ETHNOPHARMACOLOGICAL RELEVANCE: This study provides significant ethnopharmacological information, both qualitative and quantitative on medicinal plants in the Thoppampatti village, Dindigul district, Tamilnadu, India. There is urgency in recording such data to conserve the traditional medicinal plants. This is the first ethnobotanical study which records the traditional important medicinal plants of Thoppampatti village. To collect, analyze and evaluate the ethnopharmacologic knowledge in Thoppampatti village in order to protect it. This study reports the results of an ethnopharmacological survey on the uses of medicinal plants by inhabitants of the Thoppampatti. MATERIALS AND METHODS: The field study was carried out in a period of about one year (April 2012-May 2013) in Thoppampatti village. The information was obtained through open and semi-structured interviews with 48 (27 males, 21 females) knowledgeable local people and traditional healers (THs). The collected data were analyzed qualitatively and quantitatively. In addition, use value (UV), relative importance (RI) and Jaccard index (JI) was determined. RESULTS: A total of 139 species of plants, mostly trees and herbs, belonging to 54 families were identified in this study. This is used to treat 142 diseases and ailments. These ailments were categorized into 18 major categories. Leaves were the most frequently used parts, while decoction and juice are the most common method of preparation to treat various diseases. Based on IUCN Red data the identified medicinal plants include 11 least concerned species, 3 vulnerable species and one endangered species. The most important species according to their use value were Cynodon dactylon (0.79), Azadirachta indica (0.73), Ocimum tenuiflorum (0.71), Moringa oleifera (0.68), Coriandrum sativum (0.62), Abelmoschus esculentus (0.61), Acalypha indica (0.59) and Hibiscus rosa-sinensis (0.59). CONCLUSION: As a result of the present study, medicinal plants play an important role in the health care of Thoppampatti village inhabitants; they rely on medicinal plants to treat various ailments. In addition, the medicinal plants with high RI values might give some useful leads for further pharmacological investigations. Deforestation and overexploitation of a particular plant species leads to extinction so sustainable utilization of medicinal plants is recommended in the study area.

Isolation of potential probiotic Lactobacillus oris HMI68 from mother's milk with cholesterol-reducing property.

The objective of this study was to evaluate the probiotic properties of Lactobacillus strains isolated from mother's milk and their effects on cholesterol assimilation. In this study 120 isolates from mother's milk were phenotypically and genotypically characterized. Among these, only 6 predominant strains were identified as Lactobacillus spp. The following parameters were selected as important test variables in model stomach passage survival trials: acid and bile tolerance, antimicrobial activity, antibiotic susceptibility and cholesterol reduction. Results showed that the considerable variation existed among six strains. Moreover, the strain HMI68 is the most acid-tolerant and the HMI28 and HMI74 is the most acid-sensitive of all strains tested. HMI118 did not grow at 0.5% and 1% bile concentration after 5 h but the HMI68 and HMI43 showed some tolerance to such bile concentration. The differences found in the growth rate were not significant (P > 0.05). HMI68 showed resistance to most of the antibiotics as well as antagonistic activity against the tested pathogens. The amount of cholesterol reduction is increased when the media supplemented with bile salts. HMI68 assimilate 61.05 ± 0.05 µg/ml cholesterol with the presence of 0.3% bile salt this could be significantly decreased by 25.41 ± 1.09 µg/ml without bile salt. HMI68 was identified to be Lactobacillus oris HMI68 and 16S rRNA sequence was deposited in the National Center for Biotechnological Information (GenBank). For the first time the cholesterol-reducing property of L. oris isolated from mother's milk were investigated in this study. Therefore the effective L. oris HMI68 strain was regarded as a candidate probiotic.

A study on traditional medicinal plants of Uthapuram, Madurai District, Tamilnadu, South India.

OBJECTIVE: To record the medicinal plants of Uthapuram Village, Madurai district, Tamilnadu, South India for the first time and the usage of these medicinal plants to remediate the diseases among the peoples. METHODS: Explorative field trips were made to the village for about twelve months from April 2012 to May 2013 to survey the medicinal plants and collect the information from the villagers. RESULTS: From this study 52 species of valuable medicinal plants belonging to 36 families were recorded and their ethnomedicinal values were collected from the village peoples. CONCLUSION: This study focuses the importance, utilization and conservation of the medicinal plants among the people.