Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Extraction of Collagen Through Fish Waste Fermentation Optimization and Its Cytotoxic Studies on HaCaT Cell Line

Version 1 : Received: 29 September 2021 / Approved: 1 October 2021 / Online: 1 October 2021 (11:37:42 CEST)

How to cite: Sakib, S.M.N. Extraction of Collagen Through Fish Waste Fermentation Optimization and Its Cytotoxic Studies on HaCaT Cell Line. Preprints 2021, 2021100009 (doi: 10.20944/preprints202110.0009.v1). Sakib, S.M.N. Extraction of Collagen Through Fish Waste Fermentation Optimization and Its Cytotoxic Studies on HaCaT Cell Line. Preprints 2021, 2021100009 (doi: 10.20944/preprints202110.0009.v1).

Abstract

More than 800 million tons of fish are utilizing in a year and 25-30% become waste. The waste amount is beneficial source for extraction of collagen. But procedure of extraction is still to be optimized. The current study was designed to extract collagen from fish through fish waste fermentation under various conditions. For collagen extraction we did lactic acid fermentation in which yogurt and Dough bacteria was added with fish sample and placed it in incubator at 30ºC for one month. In yogurt and dough culture have at least 10 type of lactic acid bacterial species. After every week we check PH of each sample and take soup of that sample. After centrifugation of that sample’s TCA (Trichloroacetic acid) precipitation was done of yogurt and dough sample and kept at -20ºC. then did SDS-PAGE) using 6% resolving and 5% stacking gel of already stored sample. Then HaCaT cells (1 × 104 cells/well) were cultured in 96-well flat-bottom culture plates and treated with appropriate doses of FFCP (fermented fish collagen peptide) for 24 and 48 hours.The SDS-PAGE revealed that the collagen protein of fish had doublet pattern for α1 and α2 chains at corresponding to 145 kDa and 132 kDa respectively. The density for α1 twice as compared to α2. Our result agrees that The fish collagen consists mostly of α-chain as well as little amount of inter and intra molecular cross-linked components of α-chains; b (dimmer) and c (trimer). Different biochemical tests were done for identification of lactic acid bacteria catalase positive, citrate positive, urease negative. Our fermented fish collagen and peptides mixture were test for cellular cytotoxicity and proliferative effect on HaCaT cells. Current result shows that fermented extracted collagen are nontoxic and induce the proliferation of HaCaT cells. Current study is supported by various studies that revealed the medical application of fish extracted collagen as underline.

Keywords

Collagen; Environment; Fish Waste; Cytotoxic; Fermentation

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