There are several reports about the optimization of protease production, but only few have optimized the production of organic solvent tolerant keratinolytic proteases that show remarkable exploitation in the development of the non-polluting processes in biotechnological industries. The present study was carried with aim to optimize the production of a thermostable organic solvent tolerant keratinolytic protease Thermoactinomyces sp. RM4 utilizing chicken feathers. Thermoactinomyces sp. RM4 isolated from the soil sample collected from a rice mill wasteyard site near Kashipur, Uttrakhand was identified on the basis of 16S rDNA analysis. The production of organic solvent tolerant keratinolytic protease enzyme by Thermoactinomyces sp. RM4 was optimized by varying physical culture conditions such as pH (10.0), temperature (60°C), inoculum percentage (2%), feather concentration (2%) and agitation rate (2 g) for feather degradation. The result showed that Thermoactinomyces sp. RM4 potentially produces extra-cellular thermostable organic solvent tolerant keratinolytic protease in the culture medium. Further, the feather hydrolysate from keratinase production media showed plant growth promoting activity by producing indole-3-acetic acid itself. The present findings suggest that keratinolytic protease from Thermoactinomyces sp. RM4 offers enormous industrial applications due to its organic solvent tolerant property in peptide synthesis, practical role in feather degradation and potential function in plant growth promoting activity, which might be a superior candidate to keep ecosystem healthy and functional.
Plant growth promoting (PGP) rhizobacteria, a beneficial microbe colonizing plant roots, enhanced crop productivity and offers an attractive way to replace chemical fertilizers, pesticides, and supplements. The keratinous waste which comprises feathers, hairs, nails, skin and wool creates problem of solid waste management due to presence of highly recalcitrant keratin. The multi traits rhizobacteria effective to remove both keratine from the environment by producing keratinase enzyme and to eradicate the chemical fertilizer by providing different PGP activity is novel achievement. In the present study, the effective PM2 strain of PGPR was isolated from rhizospheric soil of mustard (Brassica juncea) field, Pantnagar and they were identified on the basis of different biochemical tests as belonging to Bacillus genera. Different plant growth promoting activity, feather degradation and keratinolytic activity was performed and found very effective toward all the parameters. Furthermore, the efficient strain PM2 was identified on the basis of 16s rRNA sequencing and confirmed as Bacillus cereus. The strain PM2 might be used efficiently for keratinous waste management and PGP activity. Therefore, the present study suggests that Bacillus cereus have multi traits activity which extremely useful for different PGP activity and biotechnological process involving keratin hydrolysis, feather biodegradation or in the leather industry.
Proteases have found a wide application in the several industrial processes, such as laundry detergents, protein recovery or solubilization, prion degradation, meat tenderizations, and in bating of hides and skins in leather industries. But the main hurdle in industrial application of proteases is their economical production on a large scale. The present investigation aimed to exploit the locally available inexpensive agricultural and household wastes for alkaline protease production using Thermoactinomyces sp. RS1 via solid-state fermentation (SSF) technique. The alkaline enzyme is potentially useful as an additive in commercial detergents to mitigate pollution load due to extensive use of caustic soda-based detergents. Thermoactinomyces sp. RS1 showed good protease production under SSF conditions of 55 °C, pH 9, and 50 % moisture content with potato peels as solid substrate. The presented findings revealed that crude alkaline protease produced by Thermoactinomyces sp. RS1 via SSF is of potential application in silver recovery from used X-ray films.
Free living ncmatode Caenorhabditis elegans was used as a tesi systetn for screening anthclmintic phenolics. The tnost effective concentrations (100, 500 and 1000 //g m!"') were used against root knot neinatode Meloidogyne incognita. Effect of these phenolics was determined on growth and dcveloptnent of host planr Capsicum frutescens cv. California Wonder. Second stage juveniles of M. incognita were hatched from egg tnasses collected from roots of host plant and subjected to similar phenol concentrations for 48 h. Mortality of M. incognita was recorded on the basis of parameters used for test organism bioassay. Both healthy and inoculated plants of C. frutescens cv. California Wonder were treated with solutions of salicylic acid (SA) and p-betahydroxy benzoic acid (BA) so that each pot received 100, 500 and 1000 mg phenol. Control plants were supplied with distilled water. Plants were uprooted 21 days afier inoculation and roots were gall indexed. Some plants were left in the pots ior further growth and development. Surface sterilised seedlings of host plant were raised and moculated with second stage juveniles of M. incognita. Thereafter observations were recorded on the vegetative and reproductive parameters of the plants. Drench application of SA and BA were found quite effective with no apparent phytotoxic effect. ZusammenfassungNematizidaktivitat einiger phenoiischer Verbindungen auf die Wurzelgallenbildung sowie das Wachstum und den Ertrag von Capsicum frutescens, Sorte California Wonder Die freilebende Nematode Cacnorhahditis elegans wurde als Indikator benutzt, um antihelmintische phcnolische Verbindungen zu priifen. AnschliefJend wurden die wirksamsten Konzentrationen (100, 500 und lOOO^g ml"') gegen die Wurzelgallennematode Meloidogyne incognita angewandt. Der EinfluE dieser Verbindungen auf das Wachstum und die Entwicklung der Wirtspflanze Capsicum frutescens, Sorte California Wonder wurde ermittelt. M. incognita Juvenoidtin im zweiten Entwicklungsstadium, die aus Eiermassen, gesammelt von Wurzeln der Wirtspfianze, geschllipft waren, wurden iihnlichen Phenolkonzentrationen iiber 48 h ausgesetzt. Die Sterblichkeit U.S. Copynyh, Ck.r.,KC Ccn-., Cod. St..,.mcnt: 093 1-1 785/90/2902-01 59S02.50/0
Abstract. Irradiation of isolated membranes of Sarcina lutea (Micrococcus luteus) with blue light rapidly inactivated the respiratory malate oxidase system under aerobic but not anaerobic conditions. This inactivation was much faster than that seen in whole cells suggesting that the intact organism possesses protective mechanisms capable of preventing or repairing light damage. Three photosensitive sites have been detected by comparing the effect of blue light on membranes from the carotenoid‐containing wild‐type and a carotenoidless mutant. The sites have been identified as the initial malate dehydrogenase enzyme, a flavoprotein assayed by phenazine methosulphate reduction, a sulphydryl group associated with the dehydrogenase complex but not involved in phenazine methosulphate reduction and the respiratory quinone, menaquinone. Menaquinone was found to be sensitive only in carotenoidless membranes and not in membranes from the pigmented wild‐type. Studies of the variation of photosensitivity with wavelength suggest that the three sites are sensitized by different chromophores and that the quinone acts as its own photosensitizer.
Proteases have found a wide application in several industrial processes, such as laundry detergents, protein recovery or solubilization, prion degradation, meat tenderizations, and in bating of hides and skins in leather industries. In the present study, a bacterial isolate, Thermoactinomyces sp. RS1, isolated from soil sample was taken for enzyme production by submerged fermentation technology. The classical "one-variable-at-a-time approach" was employed. Optimum incubation time, pH and temperature were found to be 24 h; pH 9.0 and 55°C, respectively. The enzyme production was highest at salt concentration of 5%; inoculum of 4% and agitation rate of 150 rpm. RS1 could grow in the presence of all carbon sources employed and was highest with glucose. In the case of organic nitrogen sources, enzyme production was highest with peptone and in the case of inorganic nitrogen sources, enzyme production was highest in urea. Overall, 1.5 folds of production was achieved after optimization of all conditions of previously used culture media. Protease in the present study shows good feather degradation within short incubation time, presenting its utilization for poultry feed production. The study holds significance as only few reports are available on the alkaline proteases having keratinolytic property from haloalkaliphilic bacteria.
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