CAJ | 학술논문

海洋假单胞菌Pseudomonas stutzeri DEH138A可以产生两种诱导型脱卤酶,即L-卤代酸脱卤酶( L-DEX)和D-卤代酸脱卤酶( D-DEX)。用DL-2-氯丙酸( DL-2-CPA)、 D-2-氯丙酸( D-2-CPA)和L-2-氯丙酸(L-2-CPA)3种诱导物分别作为种子培养与发酵培养碳源,诱导菌株DEH138A,通过HPLC检测反应体系中氯丙酸的剩余量来评估酶活性,研究底物对其酶活性的影响,并优化培养方式,实现单一构型卤代酸脱卤酶的定向诱导。结果表明： L-2-CPA可定向诱导L-DEX的产生, L-2-CPA作为唯一碳源诱导时得到的L-DEX酶活性是D-DEX酶活性的85倍,优化培养获得的L-DEX和D-DEX最高酶活性分别为6.11×10-2、3.23×10-3 U/mL (培养基),与DL-2-CPA作为诱导底物时相比较,分别提高了5.1倍和2.3倍。研究表明, L-2-CPA具有诱导专一性,实现了L-DEX的定向诱导,但对于D-DEX,则不能用L-DEX被相应构型底物诱导产生的机理解释,有待进一步研究。
해양가단포균Pseudomonas stutzeri DEH138A가이산생량충유도형탈서매,즉L-서대산탈서매( L-DEX)화D-서대산탈서매( D-DEX)。용DL-2-록병산( DL-2-CPA)、 D-2-록병산( D-2-CPA)화L-2-록병산(L-2-CPA)3충유도물분별작위충자배양여발효배양탄원,유도균주DEH138A,통과HPLC검측반응체계중록병산적잉여량래평고매활성,연구저물대기매활성적영향,병우화배양방식,실현단일구형서대산탈서매적정향유도。결과표명： L-2-CPA가정향유도L-DEX적산생, L-2-CPA작위유일탄원유도시득도적L-DEX매활성시D-DEX매활성적85배,우화배양획득적L-DEX화D-DEX최고매활성분별위6.11×10-2、3.23×10-3 U/mL (배양기),여DL-2-CPA작위유도저물시상비교,분별제고료5.1배화2.3배。연구표명, L-2-CPA구유유도전일성,실현료L-DEX적정향유도,단대우D-DEX,칙불능용L-DEX피상응구형저물유도산생적궤리해석,유대진일보연구。
The effects of carbon source substrates DL-2-chloropropionic acid( DL-2-CPA) , L-2-chloropropionic acid(L-2-CPA) and D-2-chloropropionic acid(D-2-CPA) on induction of 2-haloacid dehalogenases L-2-haloacid dehalogenase( L-DEX) and D-2-haloacid dehalogenase( D-DEX) were studied and culture process was optimized in marine bacterium Pseudomonas stutzeri DEH138A. The residual chloropropionic acid in the reaction system was quantified by HPLC. The L-2-CPA as sole carbon source was shown to specifically induce L-DEX production, with 84 times higher activity than D-DEX. The maximum activities of L-DEX (6. 11 ×10-2 U/mL media) and D-DEX (3. 23×10-3 U/mL media) were observed under culture condition of sole carbon source of L-2-CPA, 5. 1 times and 2. 3 times as the induction of DL-2-CPA. The findings indicate that L-2-CPA shows spe-cificity for directional induction of L-DEX. For D-DEX, however, it is not explained by the mechanism of L-DEX which is induced by corresponding substrate, and the induction of D-DEX needs to be further studied.