Abstract
Chitosanases and proteases have received much attention due to their wide range of applications. Four kinds of chitinous materials, squid pens, shrimp heads, demineralized shrimp shells and demineralized crab shells, were used as the sole carbon and nitrogen (C/N) source to produce chitosanases, proteases and α-glucosidase inhibitors (αGI) by four different strains of Paenibacillus. Chitosanase productivity was highest in the culture supernatants using squid pens as the sole C/N source. The maximum chitosanase activity of fermented squid pens (0.759 U/mL) was compared to that of fermented shrimp heads (0.397 U/mL), demineralized shrimp shells (0.201 U/mL) and demineralized crab shells (0.216 U/mL). A squid pen concentration of 0.5% was suitable for chitosanase, protease and αGI production via Paenibacillus sp. TKU042. Multi-purification, including ethanol precipitation and column chromatography of Macro-Prep High S as well as Macro-Prep DEAE (diethylaminoethyl), led to the isolation of Paenibacillus sp. TKU042 chitosanase and protease with molecular weights of 70 and 35 kDa, respectively. For comparison, 16 chitinolytic bacteria, including strains of Paenibacillus, were investigated for the production of chitinase, exochitinase, chitosanase, protease and αGI using two kinds of chitinous sources.
Highlights
Chitin is one of the most abundant renewable natural polymers, second only to cellulose.Chitin and its derivatives, such as chitosan and chitin/chitosan oligomers, possess great economic value because of their diverse biological activities and biotechnological applications
To reduce costs and recycle chitinous processing materials more efficiently, shrimp shells, crab shells and squid pens have been used as the sole carbon and nitrogen (C/N) sources for screening chitinolytic and proteolytic enzyme-producing bacteria [7,8,9,10]
To explore the production of chitosanase and protease via Paenibacillus, four strains (TKU029, TKU032, TKU037, and TKU042) were incubated in a liquid medium containing 1% (w/v) of different chitinous materials, which were used as the sole C/N source
Summary
Chitin is one of the most abundant renewable natural polymers, second only to cellulose. Chemical treatments, using strong alkali and acids for deproteinization and demineralization, are traditionally used to prepare chitin and chitosan from crab shells, shrimp shells and squid pens [2,3,4,5] These chemical procedures have several drawbacks, including the large amount of protein-containing wastewater produced due to the high alkali content [1]. To reduce costs and recycle chitinous processing materials more efficiently, shrimp shells, crab shells and squid pens have been used as the sole C/N sources for screening chitinolytic and proteolytic enzyme-producing bacteria [7,8,9,10]. Four chitinous materials, including squid pen powder (SPP), demineralized shrimp shell powder (deSSP), shrimp head powder (SHP) and demineralized crab shell powder (deCSP), were used as the sole C/N sources for chitosanase and protease production by Paenibacillus sp. A comparison of the α-glucosidase inhibitors produced by the four Paenibacillus strains was performed
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