ABSTRACT
A yam (Dioscorea opposita Thunb) class IV chitinase, whose genomic DNA was cloned by Mitsunaga et al. (2004), was produced by the recombinant Pichia pastoris X-33 in high yields such as 66 mg/L of culture medium. The chitinase was purified by column chromatography after Endoglycosidase H treatment and then characterized. It showed properties similar to the original chitinase E purified from the yam tuber reported by Arakane et al. (2000). This Pichia-produced chitinase also showed strong lytic activity against Fusarium oxysporum and Phytophthora nicotianae, wide pH and thermal stability, optimum activity at higher temperature such as 70 °C, and high substrate affinity, indicating that one can use this Pichia-produced yam chitinase as a bio-control agent.
Subject(s)
Biological Control Agents , Chitinases/biosynthesis , DNA, Recombinant/genetics , Dioscorea/enzymology , Dioscorea/genetics , Genetic Engineering , Pichia/genetics , Amino Acid Sequence , Chitinases/genetics , Chitinases/isolation & purification , Chitinases/pharmacology , Culture Techniques , Enzyme Stability , Fusarium/drug effects , Hydrogen-Ion Concentration , Kinetics , Phytophthora/drug effects , Pichia/growth & development , Sequence Analysis , TemperatureABSTRACT
A total of 225 carbonated soft drink (CSD) samples from nine brands, from various locations in five metropolitan cities of Bangladesh were examined to determine their bacteriological quality. Most samples were not in compliance with microbiological standards set by organizations like the World Health Organization (WHO). Pseudomonas aeruginosa was the predominant species with an incidence of 95%. Streptococcus spp. and Bacillus stearothermophilus were the next most prevalent with numbers ranging from 6 to 122 and 9 to 105 cfu/100 ml, respectively. Fifty four percent of the samples yielded Salmonella spp. at numbers ranging from 2 to 90 cfu/100 ml. Total coliform (TC) and faecal coliform (FC) counts were found in 68-100% and 76-100% of samples of individual brands, at numbers ranging from 5 to 213 and 3 to 276 cfu/100 ml, respectively. According to WHO standards 60-88% of samples from six brands and 32% and 40% of samples from two other brands belonged to the intermediate risk group with FC counts of 100-1000 cfu/100 ml. Heterotrophic plate counts, however, were under the permissible limit in all 225 samples. These findings suggest that carbonated soft drinks commercially available in Bangladesh pose substantial risks to public health.
