Optimization Of Aquatic Enzyme Conditions For Bacterial Fermentation Of Camellia Seed Oil

Aqueous enzymatic method is a new oil extraction technology studied in recent years, the waste water of Enzymatic method is most produced during the separation process, the carbohydrates, solvable nitrogen and inorganic substance it contains could be used by the microorganisms and transferred to enzyme system with the cleaning ability, combined with the tea saponin to enhance it’s cleaning function, then develop a natural detergent. The studies of the usage about waste water produced during the extraction process of Camilla oil are rare at present. This study focused on the detection of the main composition of waste water,the screening of enzyme-produced microorganisms and the optimizing of enzyme-producing conditions. The results are as follows:1.The main composition of the waste water are:saponin is about 3.6%, protein is about 5.4%,polysaccharide is 5.8%, it also contains a little amount of fat.2. The growth curve and enzyme-producing cultivation results of different kinds of microorganisms shows that bacillus subtilis has the highest yield of protease, which is 106.7 u/mL and it was chosen to be the microorganism to use the waste water. Pseudomonas aeruginosa shows the relative highest yield of lipase, which is 17.9 u/mL, so it was also chosen.3. Based on the original broth, response surface analysis shows that the best protease production condition for bacillus subtilis is:waste water addition volume 81%, carclazyte addition volume 4% and fermentation time 87 h. The protease activity is 35.9 u/mL under these condition. Verification test shows that the relative error is 1.12%.4. Response surface analysis shows that the best lipase production condition for Pseudomonas aeruginosa is:waste water addition volume 71%, carclazyte addition volume 5% and fermentation time 77h.The lipase activity is 15.6 u/mL under these condition. Verification test shows that the relative error is 1.9%.5. continuous fermentation inhibits protease production of bacillus subtilis. The later fermentation of protease activity is 27 u/mL. Presenting a decrease of 20% than prior fermentation. While later fermentation of lipase activity of pseudomonas aeruginosa is only 7.1 u/mL, presenting a sharp decrease of 46% than prior fermentation. So it is recommended to ferment pseudomonas aeruginosa first in real applications. This will help to maintain the enzymatic activities.Tea saponin addition test shows that it has no obvious linear relation inhibiting effect on bacillus subtilis. However saponin has a inhibiting effect on pseudomonas aeruginosa in the content range of 0～6.0%,with the saponin content increasing, the lipase activity decreased accordingly. There is an obvious decrease in the saponin range of 3.0%～4.5%, as the saponin content in the waste water is about 3.6%,the low lipase activity of pseudomonas aeruginosa in waste water may partly caused by this.The result above shows it is possible to develop a natural kind of detergent by the usage of waste water produced during the camellia oil extraction. Combined with the protease and lipase produced by microorganisms and under the condition of small variation of saponin content.