The health benefits associated with probiotics have increased their application in pharmaceutical formulations and functional food development. High production of probiotic biomass requires a cost-effective production method and nutrient media optimization. The biomass production of probiotics can be enhanced by optimizing growth parameters such as substrate, pH, incubation time, etc. For economical industrial production of probiotic biomass, it is required to design a new medium with low cost. Wastes from the food industries are promising components for the development of the low-cost medium. Industrial wastes such as cheese whey and corn steep liquor are excellent examples of reliable sources of nitrogen for the biomass production of probiotic bacteria. The increased yield of biomass reduced the cost of production. This review focuses on the importance of probiotic media for biomass production and its challenges. Graphical Abstract
One of the key enzymes utilized in the food industry is pullulanase. But its major drawbacks are its low yield and high production costs. In this regard, the current research aims to screen agro-waste substrates for optimal pullulanase production in solid-state fermentation. Of various agro-wastes used as a substrate, the maximum enzymic activity (9.74 U/gds) was observed in a medium based on 5 g of green gram husk and incubated for 3 days at 30 °C. The effects of 16 different nutrients on the yield of pullulanase production were studied using the Plackett–Burman experimental design. The incorporation of FeSO4, MnSO4, and MgSO4 into the pullulanase production medium significantly increased the yield and showed a 5.7-fold increase (56.25 U/gds) in comparison with the unoptimized media. The Box-Behnken experimental design was used to study the effect of interactions between Fe2+, Mg2+, and Mn2+ on the production of pullulanase. Box-Behnken showed a 1.1-fold increase (62.1 U/gds) in pullulanase production. The total increase in yield after all optimization was 6.37-fold. The present study reports for the first time the applicability of green gram husk as a potent substrate for pullulanase production by Penicillium viridicatum.
Pullulanases are the most important industrial group of enzymes in family 13 glycosyl hydrolases. They hydrolyze either α-1,6 and α-1,4 or both glycosidic bonds in pullulan as well as other carbohydrates to produce glucose, maltose, and maltotriose syrups, which have important uses in food and other related sectors. However, very less reports are available on pullulanase production from native strains because of low yield issues. In line with the increasing demands for pullulanase, it has become important to search for novel pullulanase-producing microorganisms with high yields. Moreover, high production costs and low yield are major limitations in the industrial production of pullulanase enzymes. The production cost of pullulanase by using the solid-state fermentation (SSF) process can be minimized by selecting agro-industrial waste. This review summarizes the types, sources, production strategies, and potential applications of pullulanase in different food and other related industries. Researchers should focus on fungal strains producing pullulanase for better yield and low production costs by using agro-waste. It will prove a better enzyme in different food processing industries and will surely reduce the cost of products.
For the DNA-based study of plant species, one of the important steps is to obtain high-quality DNA. However, this is problematic when the species contains a lot of polyphenols and polysaccharides. The polysaccharides and polyphenols interfere with the activity of the Taq polymerase enzyme during the PCR reaction thereby affecting the quality of the DNA. Therefore, a method for DNA extraction from Cyclanthera pedata has been developed. The current study reveals a CTAB-based approach that is quick, dependable, and economical and is specifically designed for obtaining DNA from the Cyclanthera genus. These plant species are abundant in secondary metabolites and polysaccharides, which makes it difficult to extract DNA effectively and with a high yield. The present protocol also excludes the use of expensive liquid nitrogen, which makes it cost-friendly as well. High salt concentration (1.5 M) and 2% polyvinylpyrrolidone were used in the DNA extraction buffer to prevent the solubility of polysaccharides and polyphenols in DNA extract. In addition to these substances, protein-like various enzymes were precipitated by ammonium acetate and removed by centrifugation during the isolation process. The quality of the isolated DNA was assessed using agarose gel electrophoresis (0.8%) and quantified using an A260/A280 ratio ranging from 1.7 to 1.9, absorbance ratio >2,which indicates the extract was free of proteins, polysaccharides, and polyphenols. The extracted genomic DNA was amplified by the ISSR primer (UBC-825) and clear banding pattern were observed. This standardized protocol provides pure and high quality genomic DNA without expensive liquid nitrogen or toxic phenolic compounds. It is also suitable for routine molecular biology assays such as RAPD, SSR, restriction digestion, southern blot, and cloning techniques.
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