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Mainali, P.

Publications and source records attributed to Mainali, P..

2 recordsLinked to original sources

Valorisation of spent cultivated meat media for recombinant FGF2 production in GRAS Lactococcus lactis

Innovative strategies for sustainable utilization of waste resources are imperative in the pursuit of a circular economy. Recently, the idea of utilizing mammalian spent media as a valuable resource is gaining traction, offering significant opportunities for innovative uses as a food-grade feedstock for microbial fermentation, especially in the production of alternative proteins for research and food purposes. In this study, we aim to repurpose spent mammalian culture media for production of valuable proteins. Growth factors (GFs) are a family of high-value proteins that naturally stimulate cell proliferation or differentiation. More importantly, these factors also present significant costs for cell culture. Here, we successfully demonstrate the use of spent mammalian culture media for the recombinant production of fibroblast growth factor 2 (FGF2-G3) in Lactococcus lactis. Bioreactor fermentation at a 1 L scale confirmed purified yields of 2.6 mg/L of recombinant FGF2-G3 using spent media. Further functional testing indicated that the recombinant FGF2-G3 can promote cell proliferation on an Anguilla japonica (Japanese eel) pre-adipocytic cell line, suggesting its potential for cultivated meat production. Based on the preliminary results of this study, our calculations indicate that fermenting 1 L spent mammalian waste could yield enough growth factors to efficiently grow approximately 52 L of cultivated meat through fermentation. This prediction emphasizes the potential of waste valorisation to sustainably produce protein, thus contributing to environmental preservation and economic viability. HighlightsO_LISpent mammalian culture media can be repurposed for microbial fermentation. C_LIO_LIGRAS L. lactis was used to express FGF2-G3 in an optimized spent media formulation. C_LIO_LIFunctional FGF2-G3 expression was demonstrated in controlled 1-L bioreactor runs. C_LI

microbiology↗

COMPARISONS OF CODING AND NONCODING SEQUENCES TOINFER THE ORIGIN OF CODON USAGE BIAS

Codon usage bias is the preferential use of the subset of synonymous codons during translation. In this paper, the comparisons of normalized entropy and GC content between the sequence of coding regions of Escherichia coli k12 and noncoding regions (ncRNA, rRNA) of various organisms were done to shed light on the origin of the codon usage bias.The normalized entropy of the coding regions was found significantly higher than the noncoding regions, suggesting the role of the translation process in shaping codon usage bias. Further, when the position specific GC content of both coding and noncoding regions was analyzed, the GC2 content in coding regions was lower than GC1 and GC2 while in noncoding regions, the GC1, GC2, GC3 contents were approximately equal. This discrepancy is explained by the biased mutation coupled with the presence and absence of selection pressure. The accumulation of CG content occurs in the sequences due to mutation bias in DNA repair and recombination process. In noncoding regions, the mutation is harmful and thus, selected against while due to the degeneracy of codons in coding regions, a mutation in GC3 is neutral and hence, not selected. Thus, the accumulation of GC content occurs in coding regions, and thus codon usage bias occurs.

molecular biology↗