Purification and Characterization of a Chitosanase and a Protease by Conversion of Shrimp Shell Wastes Fermented by Serratia Marcescens Subsp. Sakuensis TKU019

Abstract

A chitosanase and a protease were purified from the culture supernatant of Serratia marcescens subsp. sakuensis TKU019 with shrimp shell as the sole carbon/nitrogen source. The culture condition suitable for production of chitosanase was found to be shaken at 37 °C for 3 days in 100 mL of medium containing 0.5 % shrimp shell powder, 0.1 % K2HPO4 and 0.05 % MgSO4•7 H2O at pH 7. The TKU019 chitosanase was suppressed by the simultaneously existing protease, which also showed the maximum activity at the third day of incubation. The molecular masses of the chitosanase and protease determined by SDS‐PAGE were approximately 36 kDa and 58 kDa, respectively. Cu2+, Mn2+ and Zn2+ inhibited the chitosanase activity, and all of tested divalent metals inhibited in the protease activity. The optimum pH, optimum temperature, pH stability, and thermal stability of the chitosanase was pH 7, 60 °C, pH 4–7, and < 60 °C, respectively. The optimum pH, optimum temperature, pH stability, and thermal stability of the protease was pH 10, 50 °C, pH 5–10, and <50 °C, respectively. Tween 40 (2 %, v/v) had stimulatory effect on TKU019 protease activity.

Production and Potential Applications of Bioconversion of Chitin and Protein-Containing Fishery Byproducts into Prodigiosin: A Review

“…Chitin, chitosan, colloidal chitin, and water soluble chitosan have commonly been used as the major carbon/nitrogen (C/N) sources for the isolation of strains producing chitinolytic enzymes and the production of chitinolytic enzymes by these isolated strains [ 9 , 15 , 16 , 17 , 18 ]. To cut down fermentation expenses, the inexpensive materials of shrimp heads, shrimp shells, crab shells, and squid pens have recently been evaluated as the sole C/N sources for the production of bioactive compounds [ 5 , 6 , 9 , 12 , 13 , 14 , 15 , 16 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 ,…”

“…For example, a review of the characteristics and potential therapeutic anticancer-drug applications of PG from Serratia was introduced by Darshan and Manonmani [ 180 ], while the structure, chemical synthesis, and biosynthesis of PGs as natural products were summarized by Hu et al [ 157 ]. Additionally, synergistic inhibitory effects of chitinolytic enzymes [ 36 , 37 , 39 , 86 , 87 , 88 , 89 , 90 ], proteolytic enzymes [ 36 , 61 , 89 , 192 , 193 , 194 , 195 ], and S. marcescens PG were estimated for biocontrol in plant cultivation [ 85 , 86 , 87 , 88 , 89 , 90 ]. Biosurfactant produced by S. marcescens was also evaluated [ 167 , 196 , 197 , 198 ].…”

“…Biosurfactant produced by S. marcescens was also evaluated [ 167 , 196 , 197 , 198 ]. Recently, chitin and protein-containing marine byproducts have been utilized as the sole C/N source by S. marcescens TKU011 for the simultaneous production of PG [ 59 , 60 , 61 , 62 ], chitinolytic enzymes, and proteolytic enzymes [ 36 , 37 ]. The present review discusses the foremost accomplishments in the production and isolation of S. marcescens PG, particularly the application of shrimp heads, shrimp and crab shells, and squid pens as C/N sources for PG production by S. marcescens TKU011 [ 59 , 60 , 61 , 62 ].…”

Production of a Thermostable Chitosanase from Shrimp Heads via Paenibacillus mucilaginosus TKU032 Conversion and its Application in the Preparation of Bioactive Chitosan Oligosaccharides

“…It is a useful tool for depolymerizing chitosan into oligosaccharides with various biological activities and degrees of DP [5,9]. Important sources for chitosanase production have primarily been found in bacteria, including Paenibacillus [9,18], Bacillus [5,19,20,21,22,23], Serratia [24], and Streptomyces [25]. Until now, the common C/N sources for chitosanase production via bacteria were chitin and chitosan.…”

Bioprocessing of Squid Pens Waste into Chitosanase by Paenibacillus sp. TKU047 and Its Application in Low-Molecular Weight Chitosan Oligosaccharides Production

“…Endo-chitosanase degrades chitosan at a random position and releases COS, thus it is the major enzyme responsible for COS production [ 2 ]. Until now, numerous bacteria have been explored for their chitosanase producing ability, such as Bacillus [ 10 , 15 , 18 , 23 , 24 , 25 , 26 , 27 , 28 , 29 ], Paenibacillus [ 8 , 13 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ], Acinetobacter [ 42 , 43 ], Streptomyces [ 14 , 44 ], Serratia [ 45 ], and Pseudomonas [ 46 ]. For the production of chitosanase via bacterial fermentation, chitin is the common carbon and nitrogen (C/N) supplement.…”