Bacterial enzymes such as penicillin acylases are used for biosynthesis of betalactam antibiotics. The present study highlights the importance of manipulation of microbiological media and methods for isolation and characterization of bacterial strains having potential for penicillin acylases. Simple approach has been adopted for developing mutants with enhanced production of penicillin acylase. Bacterial strains collected from different ecological niches have unique properties and manifests diverse levels of enzyme activity. The high diversity in the natural population demonstrates that broad sampling of the bacterial population and strain improvement are necessary to select promising penicillin acylase producing bacterial strains that might be used in the industry for the synthesis of antibiotics.
Biodegradation of paranitrophenol (PNP) was carried out using Pseudomonas pseudomallei capable for PNP degradation. For enhanced biodegradation this strain was subjected to UV ray, chemical and gamma ray mutagenesis. UV and chemical mutagenesis was not fruitful as no PNP hyper degrading bacterial strain was obtained. Gamma ray mutagenesis was positive for PNP degradation and hyper degrading bacterial strain was obtained. Both parent and mutant in presence of glucose, efficiently degraded PNP. When concentration of glucose increased from 50 to 100 ppm then 50 ppm PNP was degraded within 12 hours by mutant and in 20 hours by parent. So glucose had positive effect on PNP degradation and resulted in enhanced PNP degradation. Effect of phenol was also checked on PNP degradation. The result showed that immobilization of the bacterial culture was best practice for PNP degradation. Similarly addition of glucose helped enhanced PNP degradation and ammonium sulphate provided good nitrogen source. After this toxicity of treated and untreated PNP was also checked by exposing to fish, that showed that after degradation of PNP no toxic metabolites were produced.
With the increase of world population the demand of the plant and plant products also increased tremendously since last one decade. To full fill the demand of people, plant improvement is necessary. There are various methods available for plant improvement but among those in vitro mutagenesis is a significant one.This chapter gives a detailed information about the in vitro mutation induction by various methods.
With the recent development of biotechnology, there has been vast interest to use cellulose-digestive microorganisms to convert lignocellulosic biomass to glucose that can be used in different applications such as production of fuel ethanol, use in animal feed, use in waste water treatment and in brewing industry. In the last few decades, the exponential increase in the application of cellulases in various fields demands extension in both qualitative improvement and quantitative enhancement. Quantitative enhancement requires strain improvement and medium optimization for the overproduction of the enzyme as the quantities produced by wild strains are usually too low. The spectacular successful examples of strain improvement in industry are mostly attributed to the extensive application of mutation and selection. Such improved strains can reduce the cost of the processes with increased productivity and may also possess some specialized desirable characteristics. Thus, enhanced cellulase production by strain improvement of Bacillus PC-BC6, optimization of process parameters and using pretreated substrate can be a cheaper source.
Enzyme is a protein biocatalyst produced by living cells. They bring about & control the numerous biochemical reactions involved in metabolic process of cells.They are widely applied in research, industry & diagnostic purposes. In this book production of such commercially & diagnostically important enzyme Glucose Oxidase is discussed whichbelongs to the class-oxidoreductase. Glucose oxidasehas a large demand all over the world. Currently it is produced from Asp.niger &Penicillium notatum. In our laboratory bacterial source of this enzyme is isolated, which is Pseudomonas aeruginosa from paper mill waste. The production at laboratory scalehas been standardized. The strain is deposited atNational Collection Ind.& Marine Bacteria (NCIMB) Abberdeen U.K.with accession number 13118. The strain was mutated with UV-rays &EMS. The mutant strain showed 40 percent enzyme activity extracellular. The other characters were also studied. This is a novel bacterial source of glucose oxidase.
Phosphate solubilizing microorganisms were isolated from rhizosphere soils of rice fields Tiruchirappali district of Tamilnadu. The isolated cultures were identified as Pseudomonas sp, Aspergillus niger, Aspergillus fumigatus and Penicillium sp. The efficacy of phosphate solubilization, Phosphatase, Lipase activity and Indole acetic acid production assay of isolated strains were carried out. The isolated strains were subjected to random mutational studies. The phosphate solubilization, Phosphatase, Lipase activity and Indole acid production were carried out after random mutagenesis by using wild and mutated cultures. Phosphate solubilization efficacy of the wild strains was higher in fungi when compared to bacteria. The phosphate solubilization efficacy and Phosphatase activity of the wild strains was observed in the order of A.niger >Penicillium sp. > A.fumigatus. ANems120 showed the highest phosphate solubilization activity among all chemical mutated fungal strains.ANsa120 was the predominant fungal strain for Phosphatase activity. The PSems120 was the predominant Lipase producing strain. All the strains except A.fumigatus have the ability to produce the plant hormone IAA.
Citric acid is a tri carboxylic organic acid is the most important acid produced by fermentation used in the food and pharmaceutical industries. The present work deals with determination of optimum growth conditions for citric acid production by submerged fermentation by using strain of A.niger MTCC662 respectively. Isolation, screening and mutated the A.niger strain induced by UV irradiation, diethylsulphate and Co60 by providing different types of media and the submerged fermentation was carried out under various growth parameters like temperature, pH and media of Sorghum malt as substrate. Strain improvement studies for maximum production of citric acid obtained after the fermentation. By using the response surface methodology has been optimized the production of citric acid. These variables were further optimized using a 24 full factorial CCD (Central Composite Design) and a second order polynomial model equation was obtained.
Bacterial wilt caused by Pseudomonas solanacearum is a serious plant disease in tropics and warm climate regions in the world. Agrobacterium mediated transformation was used for introduction of Xa21 gene for enhanced resistance to bacterial wilt in tomato cultivars. Factors effecting transformation such as pre-selection, preculturing of explants with acetosyringone had improved transformation efficiency. There are very few reports addressing the molecular mechanism of the plant responses to bacterial attack. Tomato cultivars were differentiated on the basis of their responses to bacterial wilt strain in to resistant and susceptible cultivars. Proteomic profiling was done for understanding of bacterial defenses. Molecular chaperones such as HSP70, 60 kDa chaperonin, protein disulfide isomerase & apical membrane antigen are speculated to have role in plant defenses against bacterial infection. Peroxiredoxin, ATP synthase, S-adenosyl methionine synthase, arginase and iron ABC transporter are other candidate proteins speculated for plant defense via JA and SA pathway.
Escalating trends in fossil fuel consumption has lead to shortage of energy domains. Cellulosic biomass is regarded the most promising substitute for supplementation of energy demands. Enzyme production from lignocellulosic biomass via biological route seems to be very attractive and sustainable due to ubiquitous nature of biomass and its non-competitiveness with food crops. The key element in bioconversion process of lignocellulosics to biofuels is the hydrolytic enzymes chiefly cellulases. Cellulases are the third largest industrial enzyme in the world and gaining rejuvenated interest due to tremendous applications. Therefore, reduction in cellulase production cost, improvement in cellulolytic potential of microbes and enhancement in sugar yields are extremely accountable to bioethanol production cost. In this context, improvement of microbial strains for over expression of cellulase has evoked tremendous research efforts in commercial fermentation process. Conclusively, proficient cellulase yield by strain improvement of Trichoderma viride and process optimization by exploiting cheaper substrate can govern cost reduction in enzyme production process.
Four bacterial cytochrome P450 enzyme activities are described in terms of their potential biotechnological use for steroid transformations, by engineered ''Rhodococcus'' strains, into commercially valuable compounds. Three selected rhodococcal P450s were characterized with microbiological, genetic, biochemical, spectroscopic and chromatographical methods. Two sterol C26- hydroxylases were demonstrated to be important for 3- hydroxysterol degradation. The role of the novel dextromethorphan N-demethylase in sterol metabolism was discussed. Furthermore, heterologous production of a mutated P450 enzyme, from ''Bacillus megaterium'', was shown in an engineered ''Rhodococcus'' strain. The activity of the mutated P450 was identified as 16?-hydroxylase of 4- androstene-3,17-dione, by ?H and ??C NMR. The detailed characterization of the cytochrome P450 enzymes, described in this book, provides insight into the catabolic repertoire of selected bacterial strains, necessary for engineering of steroid catabolic pathways in order to produce pharmaceuticals efficiently.
Chemical Mutagenesis in Chicory: A Tool For Crop Improvement Chicory is a well known medicinal herb used for treatment of liver ailments. The herb grows wild and it still has low genetic variability being self pollinated. This opens an ample room for mutation breeders to improve the crop. Mutation in plants for improvement of potential agronomic traits is a buzz word of today and had become one of the most important tools in generating new varieties. Mutation breeding employing varieties of physical and chemical agents is in use to explore the possibilities of developing new varieties, especially in crops having narrow genetic base. The present study describes the effect of chemical mutagens in induction of mutations in Cichorium intybus (L.) since genotype of the plant is homozygous with limited genetic variability, hence induced mutagenesis proved to be an effective tool in induction of mutations and further improvement in the crop via selection.
The isolated bacterial strains of Planomicrobium chinense, Bacillus cereus, Pseudomonas aeruginosa and Klebsiella oxytoca from central India are implied to be potential degraders of petroleum hydrocarbons. Diesel degrading and IAA producing strain of P. chinense may be used as a promising plant growth promoter under xenobiotic stress conditions after strain improvement. Such findings are likely to provide an insight into the exploitation of such microorganisms for degradation of diesel oil spills in tropical environment. Further studies on characterizing the genes of the four strains is likely to throw more light on the molecular mechanism underlying the process of bioremediation.
Prodigiosin was isolated from fermented broth of isolated marine bacterial strain Vibrio sp. The compound was characterized, and evaluated for its antimicrobial properties. The isolated strain was molecularly identified based on 16S rRNA based phylogenetic analysis. This molecular analysis data suggested that this strain is closely related to Vibrio sp. The extract was purified and characterized by spectral data using NMR and MASS and confirmed as analogue 2,2'-[3-methoxy-1'amyl-5'-methyl-4-(1''-pyrryl)] Prodigiosin structure. Prodigiosin exhibited antioxidant and broad spectrum antimicrobial properties against gram negative and gram positive strains. Molecular docking study with different cancer targets revealed that this compound it may be a potential candidate at pharmaceutical sector.
A series of novel 2-(substituted benzylidene)-7-(4-bromophenyl)-5-(furan-2-yl)-2H-thiazolo[3,2-a]pyrimidin-3(7H)-ones were synthesized by using established methods and were characterized by IR, 1H NMR and Mass spectroscopy in order to confirm their formation. These compounds were synthesized to evaluate their antitubercular, antibacterial and antifungal potency. Docking studies revealed the importance of functional groups and its different biological action. They were found to be potent against Mycobacterium tuberculosis H37Ra strain, also potent against Gram +ve, Gram –ve and fungal strains
A strain of egg parasitoid, Trichogramma chilonis was developed for tolerance to insecticides and high temperature. The LC50 values were 13-folds higher for endosulfan, 233-folds for monocrotophos and 4- folds for fenvalerate. In high temperature, the developed strain could parasitise up to 53.3 eggs as compared to 6.7% by susceptible strain. Exact identification of developed strain was done using primers, viz., OPA-4,OPC-12 and OPL-4. Increased GST activity was recorded in developed strain, whereas decrease in carboxylestrase and similar acetylcholine activity to susceptible strain. Genetical studies with tolerant strain of T. chilonis revealed that female parent provide incomplete dominant characters and male provide recessive dominance. Partial sequence of Heat shock protein 70 (hsp70) was done for tolerant strain. Net house studies conducted on cotton, tomato and cabbage revealed that developed strain parasitised eggs to the tune of 60.0-70.0% as compared to 5.0- 15.0% by a susceptible strain. Field trials carried out on cotton, rice, tomato, cabbage, brinjal, okra and sugarcane proving efficacy of developed strain all over India.