ABSTRACT
A thermophilic Geobacillus bacterium secreting high activity of endo-glucanase (EC 3.2.1.4) was isolated from rice straw compost supplemented with pig manure. A full-length gene of 1,104 bp, celA, encoding this glycosyl hydrolase family 5 endo-glucanase of 368 amino acids was isolated. No related gene from Geobacillus has been reported previously. The recombinant CelA expressed in Escherichia coli had an optimal activity at 65 degrees C and pH 5.0, and it exhibited tenfold greater specific activity than the commercially available Trichoderma reesei endo-glucanase. CelA displayed activity over a broad temperature range from 45 to 75 degrees C and was a thermostable enzyme with 90% activity retained after heating at 65 degrees C for 6 h. Interestingly, CelA activity could be enhanced by 100% in the presence of 2 mM MnSO(4). CelA had high specific activity over beta-D-glucan from barley and Lichenan, making it a potentially useful enzyme in biofuel and food industries.
Subject(s)
Bacillus/enzymology , Glycoside Hydrolases/isolation & purification , Amino Acid Sequence , Bacillus/genetics , Base Sequence , DNA Primers , DNA, Ribosomal/genetics , Enzyme Stability , Escherichia coli/genetics , Glycoside Hydrolases/genetics , Glycoside Hydrolases/metabolism , Hydrogen-Ion Concentration , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Substrate Specificity , TemperatureABSTRACT
Replicated field trials at three matched farm pairs in southern and central Taiwan were established in October 2004 and 2005 to compare fruit quality and nutritional parameters of tomatoes grown on-farm under organic versus conventional management systems in tropical and subtropical environments. Two processing tomato varieties were evaluated using a randomized complete block design at each of the farms. Aggregation of farms by type (organic vs conventional) across two years resulted in no significant differences between organic and conventional farming systems for all tomato fruit parameters measured, including quality (pH, soluble solids, acidity, and color), content of bioactive compounds with antioxidant activity (beta-carotene, lycopene, ascorbic acid, and total phenolics), and antioxidant activity. This study indicated no consistent effect of the farming system on tomato fruit parameters. Farm management skills combined with site-specific effects contributed to high lycopene levels, and the choice of variety significantly influenced the content of bioactive compounds, particularly ascorbic acid and total phenolics.
