Study On The Production Of Pancreatic Lipase Inhibitor Of Microbial Screening,Identification And Fermentation Conditions

Obesity was an important public health and medical problems becoming one of the top ten threats to human health that confirmed by a spokesman of the World Health Organization?WHO?.Currently,there are more than one billion adults heavier than normal all over the world.The pancreatic lipase inhibitor is a new type of diet pills that has the advantage of relative safety.It doesn't act on the nervous system and blood circulation system.It inhibited the absorption of food in the stomach.For this study,we collected a large number of soil samples,and screened them by improved plate method.We want to find some microbial that can produce pancreatic lipase inhibitor.We also used UV-visible spectrophotometer for rapid detecting fermentation broth of microbial.Then we found a fermentation broth containing lipase active ingredient and purify strain.The morphological character of colony,physiological and biochemical characteristic,16SrDNA sequence analysis and phylogenetic tree of the strain were observed,we identified it as Bacillus subtilis and named it as Bacillus subtilis z123.We used single-factor test method for optimum fermentation condition and optimum medium composition of strain.We got a media component containing 15 g/L of malt extract as carbon source,8g/L of yeast extract as nitrogen source,5 g/L NaCl.Also a small amount of Fe²⁺?Ca²⁺?Mg²⁺metal ions were good to cultivating strain.The initial value--pH7.6,the optimum culture temperature--30?,the optimum inoculum--4%,were the best fermentation conditions.We prepared immobilized lipases and used them to separate for pancreatic lipase inhibitor and got immobilized enzyme inhibitor combination products.Add the said substance into 3mL of ultrapure water and boil water bath for 90s,then the lipase lose activity and release inhibitors.Centrifugation the water,we got a result of the supernatant with 22%inhibitory activity.And HPLC analysis contained the peak time of 2.344 s.We considered it maybe a new pancreatic lipase inhibitor ingredient.