Conversion of Squid Pens to Chitosanases and Proteases via Paenibacillus sp. TKU042

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.

Conversion of Shrimp Head Waste for Production of a Thermotolerant, Detergent-Stable, Alkaline Protease by Paenibacillus sp.

“…Every year, the seafood processing industry discards a large amount of by-products, including viscera, shells, heads, squid pens, fins, and bones, even though they could be recycled to produce bioactive compounds like gelatin [1][2][3][4], enzymes [4][5][6][7][8][9][10][11][12][13][14][15][16][17], chitin [8,[18][19][20][21][22][23][24][25][26], chitin oligomers [7,11,12], α-glucosidase inhibitors (aGI) [27][28][29][30][31], carotenoids [32,33], and bioactive peptides [34][35][36][37][38][39][40]. Consequently, much research has gone into converting these by-products into bioactive products that have potential applications in biotechnological, agricultural, nutritional, pharmaceutical, and biomedical industries [1,…”

“…However, there are several drawbacks to the use of chemical procedures. With a more environmentally friendly production process, these marine byproducts could be used as carbon/nitrogen sources for microorganism bioconversion to various high-value products, including chitinase/chitosanase [6,[11][12][13]15,16,20], proteases [13,14], exopolysaccharides [41][42][43], and tyrosinase inhibitors [44].…”

“…Paenibacillus is a useful bacterial family employed in the medical, industrial, and agricultural fields [48]. Recently, when fishery processing by-products were used as the nutrition source (carbon and nitrogen), several strains of Paenibacillus exhibited excellent abilities in the production of bioactive compounds, including α-glucosidase inhibitors [49], exopolysaccharides [43], anti-inflammatory medication [50,51], antioxidants [30], and chitosanases [11][12][13]. However, few reports have examined proteases derived from Paenibacillus, especially using shrimp heads.…”

“…However, few reports have examined proteases derived from Paenibacillus, especially using shrimp heads. While several Paenibacillus strains demonstrated protease productivity on media containing squid pen or demineralized crab shell, the properties of their proteases were not explored [13].…”

Coagulation of Chitin Production Wastewater from Shrimp Scraps with By-Product Chitosan and Chemical Coagulants

“…As a consequence, the industry produces large quantities of wastewater streams that contains a significant number of chemical residues with highly acidic and highly alkaline flows, as well as high levels of organic matter (mostly protein), minerals, and pigments that are toxic to the environment and a risk to human health . Currently, chitin production processes have not widely adopted the use of cleaner production methods that can increase the efficiencies of raw material utilization and reduce the environmental pollution issues [5,[7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25]. In addition, the key organic were taken from the equalization tank of the wastewater treatment system and characterized as shown in Table 1.…”