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Production, Purification, and Application of the Microbial Enzymes
Published in Pankaj Bhatt, Industrial Applications of Microbial Enzymes, 2023
Anupam Pandey, Ankita H. Tripathi, Priyanka H. Tripathi
Enzymes having a role in the various industrial application are usually isolated from several normal to extreme environments. Novel microbial enzymes have been discovered using high-throughput screening technologies; however, the problem is that these bacteria are difficult to cultivate in the lab and produce a low amount of enzyme. As a result, companies have used rDNA technology to boost the yield of specific enzymes by cloning the enzyme coding gene and using heterologous expression. Insulin, for example, has been produced using rDNA technology. The genetically altered E. coli produces 100 times more insulin than native microbial strains, making insulin more readily available, at a low cost, and in large quantities (Chiang, 2004). As a result, industries are turning to genetically modified microbial strains to produce enzymes like lipases and amylases, which have a wide range of applications in the food industry (Olempska-Beer et al., 2006).
Development of Industrial Strain, Medium Characteristics and Biochemical Pathways
Published in Debabrata Das, Soumya Pandit, Industrial Biotechnology, 2021
Recombination: When two different genetical strains combine to generate a hybrid that is superior and different from either of the parents. Recombination is useful in erasing the neutral and deleterious results which arise during random mutagenesis. Recombinant DNA technology helps in improvement of primary metabolites such as amino acids and extracellular enzymes (Figure 2.10). Recombination techniques involve cloning of genes and expression of the cloned gene into an expression vector and its product formation. Isolation and cloning of genes of interest are the important aspects in rDNA technology which helps in constructing important enzymes and transfers the genes into a suitable host organism. Production of recombinant proteins and metabolic engineering are the two important objectives of recombinant DNA technology (Rowlands, 1984b).
DNA Structure, Sequencing, Synthesis, and Modification: Making Biology Molecular
Published in Richard J. Sundberg, The Chemical Century, 2017
Recombinant DNA (rDNA) refers to DNA that contains segments from two or more sources. The resulting DNA is sometimes called a chimera, referring to mythological creatures that were combinations of two different animals. Efficient and selective recombination was reported in 1973, and the key to success was a restriction endonuclease, EcoRI, that had the property of asymmetric cleavage of DNA, thus leaving cohesive (“sticky”) termini that could be reconnected. DNA recombination was known to occur in nature and small amounts of incorporation had been noted by other methods, but the recombination based on EcoRI was much more efficient.
Impact of inoculum acclimation on energy recovery and investigation of microbial community changes during anaerobic digestion of the chicken manure
Published in Environmental Technology, 2020
Cigdem Yangin-Gomec, Tugba Sapmaz, Sevcan Aydin
On the other hand, microbial quantification results using 16S rDNA-based qPCR assay showed that bacteria, archaea and methanogen amounts increased in both reactors as the operation continued. When microbial results were compared; analysis of the methanogenic community showed limited differences between the two reactors whereas similar amounts of archaeal cells were quantified in the biomass samples taken from the unadapted and adapted reactors by the end of about two-month operation. According to the NGS results of the samples from the unadapted bioreactor after about three more months; the Firmicutes and Bacteroidetes bacteria as well as methanogenic archaea-related communities in the anaerobic digesters played a functional role for anaerobic degradation of chicken waste. Hence, failure to maintain the stability of these phyla might cause a decrease in the efficiency of anaerobic bioreactors digesting chicken manure.
Bacterial and archeal dynamics of a labscale HYBRID gas fermentation bioreactor fed with CO2 and H2
Published in Journal of Environmental Science and Health, Part A, 2019
Burcin Karabey, Sidika Tugce Daglioglu, Nuri Azbar, Guven Ozdemir
VFA concentration and biomethane production results, that effected with different concentrations of H2: CO2, are quite related to the shiftings on microbial diversity. The 16s rDNA genes were amplified from the total DNA extracted from nine reactor samples with bacteria- and archaea-specific primers, respectively. Sample A represents anaerobic granular seed sludge and is also the source of the initial microbial community. The microbial diversity of sample A (seed sludge) is of great importance, as it is directly related to the methane production potential to be obtained during operation. Sample B (Filtrate) and C (Granular sludge) were also investigated since they represent the microbial diversity of the reactor that methane production rate is relatively high as 5.4 m3 CH4/m3 reactor/d at H2: CO2 ratio of 4:1.
Molecular biological tools in concrete biodeterioration – a mini review
Published in Environmental Technology, 2019
Vinita Vishwakarma, Balakrishnan Anandkumar
The presence of functional genes in concrete sewer biofilms was determined using published PCR assays [90]. Specifically, the following genes were screened: ammonia monooxygenase [91], adeno-5′-phosphosulfate reductase [92], catalytic subunit of nitrous oxide reductase [93], NiFe hydrogenase [94], Geobacteraceae specific SSU rDNA [95], aerobic methanotrophs [96], sulphur-oxidizing subunit B [97], type I methanotrophic bacteria SSU rDNA [96], type II methanotrophic bacteria SSU rDNA (McDonald et al. 2008), archaea SSU rDNA [98], general ITS fungal primers [98], sulphite reductase [99] and Shewanella SSU rDNA [100]. Quantification and identification of specific microorganism by using primers and quantitative PCR represents the presence of specific microbial group or species qualitatively and quantitatively in the concrete biofilms.