Mannitol

OFAT method was carried out by changing parameters of factors in a sequential manner. The factors considered for optimization were pH of the media, Incubation temperature, Carbon and nitrogen sources and presence of metal ions. The pH of the media was adjusted between 4 and 10. The incubation temperature ranged between 4 °C and 45 °C. Additional carbon sources like glucose, fructose, sucrose, dextrose, inositol, mannitol, rhamnose, maltose, lactose, trehalose, sodium citrate, arabinose (HiMedia Laboratories, Mumbai, India, LR grade) was supplemented at 10 g/L along with Starch. The effect of nitrogen source on amylase production was evaluated by replacing peptone with malt extract, beef extract, yeast extract, peptone, casitone, corn steep liquor (HiMedia Laboratories, Mumbai, India, LR grade) and inorganic nitrogen sources such as ammonium sulphate, ammonium chloride, ammonium carbonate, potassium nitrate and sodium nitrate (HiMedia Laboratories, Mumbai, India, LR grade). Various metal ions such as Fe²⁺, Cu²⁺, Mg²⁺, K⁺, Ca²⁺, Mn²⁺, Ba²⁺, Sn²⁺, Zn²⁺, Co²⁺, Ni²⁺at 0.5 g/L were supplemented in the media in the form of metal salts. All the experiments were performed in triplicates and the data is represented in Mean ± Standard deviation.

Silymarin and Ch were generously gifted by Micro Labs, Bengaluru (India) and Central Institute of Fisheries and Technology, Kochi (India) respectively. Sodium tripolyphosphate (TPP) and mannitol were procured from Himedia laboratories Pvt. Ltd., Mumbai (India). All other chemicals were of analytical grade and were used as and when supplied.