Pestalotiopsis sp. J63, producing a high activity of laccase, is a new marine-derived fungus isolated from the oceanic sediment of the East China Sea. Since the marine environment is oligotrophic nutrient, marine deri...Pestalotiopsis sp. J63, producing a high activity of laccase, is a new marine-derived fungus isolated from the oceanic sediment of the East China Sea. Since the marine environment is oligotrophic nutrient, marine derived fungi may use small amount of nutrients to grow and produce laccases. Agricultural residues that are mainly composed of lignin, cellulose and hemicellulose are difficult to be degraded and few microbes can take them as substrates, so they are considered as oligotrophic nutrient and have the potential to be used to produce value added products. In this study, the ability of Pestalotiopsis sp. J63 to use agricultural residues to produce laccases was tested in the submerged fermentation. The combination of 3 g·L 1maltose and 20 g·L 1rice straw was the best carbon sources and 8 g·L 1ammonium sulfate was the best nitrogen source under the condition without inducers. The effects of five inducers, the feeding time and concentration of inducer on laccase production were investigated.Adding 0.09 mmol·L 1phenol after 24 h of incubation led to high laccase activity(5089 U·L 1), while with 0.09mmol·L 1phenol in the medium and wheat bran as the nitrogen source, the laccase activity could reach 5791.7U·L 1. Native-PAGE results showed that two laccase isozymes were present in the cultures. One existed in both induced and non-induced culture filtrates, while the other was only found in the fermentation with the addition of phenol, guaiacol and veratryl alcohol.展开更多
Laccase was immobilized on the ceramic-chitosan composite support by using glutaraldehyde as the cross-linking reagent. The immobilization conditions and characterization of the immobilized enzyme were investigated. T...Laccase was immobilized on the ceramic-chitosan composite support by using glutaraldehyde as the cross-linking reagent. The immobilization conditions and characterization of the immobilized enzyme were investigated. The immobilization of laccase was successfully realized when 3.0 mL of 1.25 mg/mL of laccase at a pH value of 4.0 reacted with 0.15 g of ceramic-chitosan composite support(CCCS) at 4 ℃ for 24 h. The immobilized enzyme exhibited a maximum activity at pH 3.0. The optimal temperatures for immobilized enzyme were 25 ℃ and 50 ℃. The K_m value of immobilized laccase for ABTS was 66.64 μmol/L at a pH value of 3.0 at 25 ℃. Compared with free laccase, the thermal, operating and storage stability of immobilized laccase was improved after the immobilization.展开更多
基金Supported by the National Natural Science Foundation of China （21036005） and Scientific and Technology Plan of Zhejiang Province （2011C33016）.
文摘Pestalotiopsis sp. J63, producing a high activity of laccase, is a new marine-derived fungus isolated from the oceanic sediment of the East China Sea. Since the marine environment is oligotrophic nutrient, marine derived fungi may use small amount of nutrients to grow and produce laccases. Agricultural residues that are mainly composed of lignin, cellulose and hemicellulose are difficult to be degraded and few microbes can take them as substrates, so they are considered as oligotrophic nutrient and have the potential to be used to produce value added products. In this study, the ability of Pestalotiopsis sp. J63 to use agricultural residues to produce laccases was tested in the submerged fermentation. The combination of 3 g·L 1maltose and 20 g·L 1rice straw was the best carbon sources and 8 g·L 1ammonium sulfate was the best nitrogen source under the condition without inducers. The effects of five inducers, the feeding time and concentration of inducer on laccase production were investigated.Adding 0.09 mmol·L 1phenol after 24 h of incubation led to high laccase activity(5089 U·L 1), while with 0.09mmol·L 1phenol in the medium and wheat bran as the nitrogen source, the laccase activity could reach 5791.7U·L 1. Native-PAGE results showed that two laccase isozymes were present in the cultures. One existed in both induced and non-induced culture filtrates, while the other was only found in the fermentation with the addition of phenol, guaiacol and veratryl alcohol.
文摘Laccase was immobilized on the ceramic-chitosan composite support by using glutaraldehyde as the cross-linking reagent. The immobilization conditions and characterization of the immobilized enzyme were investigated. The immobilization of laccase was successfully realized when 3.0 mL of 1.25 mg/mL of laccase at a pH value of 4.0 reacted with 0.15 g of ceramic-chitosan composite support(CCCS) at 4 ℃ for 24 h. The immobilized enzyme exhibited a maximum activity at pH 3.0. The optimal temperatures for immobilized enzyme were 25 ℃ and 50 ℃. The K_m value of immobilized laccase for ABTS was 66.64 μmol/L at a pH value of 3.0 at 25 ℃. Compared with free laccase, the thermal, operating and storage stability of immobilized laccase was improved after the immobilization.
