Prearation And Application Research Of The Penicillinase

β-lactalnases are enzymes produced by some bacteria and are responsible for their resistance toβ-lactam antibiotics.β-lactmases are clsaafied into A,B.C and D groups according to their structure and function.Penicillinase belongs to class A group and it can hydrolyze penicillin,cephalosporins.Oximeβ-lactams,cloxacillin,carbenicillin. carbapenems antibiotics,and its activity can be inhibited bv clavulanic acid.Penicillinases are mainly used in sterile testing for antibiotic drug and also used for biosensors and antibody for ELISA.There is no report or industried-grade products.The main results of this paper are as follows:Firstly,Bacillus cereus kd-02,a high-yielding penicillinase bacteria,was obtained under anxanography method by UV mutagenesis to Bacillus cereus kd-01.The kd-02 strain’s fermentation broth concentration reached to 3 016 000 U/mL.Secondly,the fermentation processs was optimized.The penicillinase fermentation concentration reached to 7 588 000 U/mL,on the average of 7 500 000 U/mL.The optimized conditions were that Bacillus cereus kd-02 was cultivated in 50 mL/250 mL liquid fermentation medium(pH7.0～7.2) with 10%inoculum at 230 r/min,30℃.Thirdly,the preparation process for steril peniciliinase was obtained by 0.22μm filter membrane with 92.3%efficiency.Fourthly,the penicillinase stability was investigated and the result showed that the enzyme can store at below 25℃for a month with more than 92%activity.Lastly,the sterile penicillinase was surveyed for its application on penicillin slag.The 2 179 U/g(about 1.3 mg/g) penicillin was completely hydrolyzed by our penicillinase preparation at 37℃,lhr.The optimized the penicillinase and penicillin ratio was 3:1.