Factors influencing activity of enzymes and their kinetics: bioprocessing of skin.
The conventional chemically based method of dehairing and fiber-opening discharges an enormous amount of pollutants in the processing of skins. Hence, bioprocessing of skin through a two-step process, dehairing using protease and fiber opening using alpha-amylase, has been developed. However, because this process involves two steps, we characterized commercial protease and alpha-amylase for their optimum activity and determine the influence of one enzyme on the activity of the other, in order to develop an integrated enzymatic dehairing and fiber-opening process. The influence of various factors, substrate concentration, time, pH, and temperature, on the activity of both protease and alpha-amylase was determined. Furthermore, the activity of protease on mixing with alpha-amylase and vice versa was investigated. It was found that there was no significant change in the activity of one enzyme in the presence of the other. Lineweaver-Burk plots showed K(m) and V(max) values of 31.6 mg/mL and 0.0106 mg/(mL@min) for protease and 8.79 mg/mL and 0.0912 mg/(mL@min) for alpha-amylase. This study provides substantial evidence for integrating the enzyme-based dehairing and fiber-opening processes using both the selected protease and alpha-amylase in one step.
Madhumathi
M
M
Chemical Laboratory, Central Leather Research Institute, Adyar, Chennai 600020, India.
Cheerla
Srinija
S
Saravanabhavan
S
S
Thanikaivelan
P
P
Rao
J Raghava
JR
Babu
N K Chandra
NK
Nair
Balachandran Unni
BU
D000818:Animals; D002417:Cattle; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010447:Peptide Hydrolases / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D012867:Skin / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors; D000516:alpha-Amylases / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*
D000516:alpha-Amylases; D010447:Peptide Hydrolases