An affinity ligand was designed from 1-aminocyclohexane based on the crystal structure of Streptomyces albidoflavus phospholipase A₁ (saPLA₁) by using Discovery Studio software. The molecular docking results indicated that the designed ligand could interact with the active pocket of saPLA₁. Epichlorohydrin, cyanuric chloride and 1-aminocyclohexane were used to synthesize the affinity ligand, which was composed to Sepharose beads. The density of the ligand on Sepharose beads was 22.5 ± 1.1 μmol/g wet gel. Adsorption analysis of the sorbent indicated the maximum adsorption (Q_max) of the enzyme was 10.7 ± 0.29 mg/g and the desorption constant (K_d) was 426.6 ± 29.7 μg/mL. The sorbent could bind the enzyme in the supernatant of disrupted recombinant Escherichia coli through one step of affinity adsorption. After the optimization of the purification process, a single band was obtained at approximately 30 kDa, which was confirmed as saPLA₁ by the matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry and activity assay. The purity of the isolated enzyme was about 96.6 % with the purify fold at 7.62, and the activity recovery was 52.5 %.

使用Discovery Studio软件，根据链霉链霉菌磷脂酶A ₁（saPLA ₁）的晶体结构，从1-氨基环己烷设计亲和配体。分子对接结果表明，设计的配体可以与saPLA ₁的活性口袋相互作用。用环氧氯丙烷，氰尿酰氯和1-氨基环己烷合成亲和配体，该配体由琼脂糖珠组成。Sepharose珠上的配体密度为22.5±1.1μmol/ g湿凝胶。吸附剂的吸附分析表明该酶的最大吸附（Q _max）为10.7±0.29 mg / g，解吸常数（K _d）为426.6±29.7μg/ mL。通过一步亲和吸附，吸附剂可以与被破坏的重组大肠杆菌上清液中的酶结合。优化纯化过程后，在约30 kDa处获得一条条带，通过基质辅助激光解吸/电离串联飞行时间（MALDI-TOF / TOF）质谱和活性确认为saPLA ₁分析。分离的酶的纯度为约96.6％，纯化倍数为7.62，活性回收率为52.5％。