Judicial interpretation of consequential loss in construction contract

Abstract

The chitinase production by Trichoderma virens using shrimp waste as a substrate was studied in solid state fermentation with 70% of moisture content. Six different pretreatment methods namely oven pretreatment, microwave pretreatment, boiling and crushing pretreatment, sun-dried pretreatment and chemical pretreatment were conducted on shrimp waste with non-treated shrimp waste as a control. The highest chitinase activity was obtained from microwave pretreatment on the third day of fermentation with chitinase activity of 0.194 U/g IDS, 3.2 fold higher than the untreated shrimp waste (0.06 U/g IDS). Study on the effect of nitrogen source on chitinase production using general factorial design showed that ammonium sulphate with 30.29 mM nitrogen gave significant effect compared toyeast extract with 7.43 mM nitrogen. Two level factorial design, incubation time, temperature, and substrate moisture also have a significant impact on the production of chitinase. Central composite design (CCD) was used in optimizing the conditions for chitinase production of shrimp waste by solid-state fermentation. Chitinase production was found to have increased 2.46 times (0.487 U/g IDS) at optimum condition: temperature of 27.9 ° C, 54.5% of substrate moisture and six days of incubation time. The optimal degradation showed an improvement of chitinase production of 2.46 fold as compared to before optimization using CCD. For partial characterization of chitinase, the optimum temperature and pH are at 60 °C and pH 3.0, respectively. Chitinase retained 72% of its activity at 70 °C. However, the loss of the chitinase activity occurred after 60 minutes of incubation at 70 °C and 80 °C with residual activity are 48% and 28%, respectively. Chitinase was more stable in acidic than in alkaline pH. The molecular weight of chitinase was 50 and 42 kDa for endochitinase, 33 and 25 kDa for eksokitinase and 18 kDa for protease. Extraction of crude chitinase from Trichoderma virens can inhibit the growth of Ganoderma boninense.