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目的探讨罗非鱼皮酶溶性胶原蛋白(Pepsin soluble collagen peptide from Tilapia skin,PSCPT)、罗非鱼皮酸溶性胶原蛋白(Acid soluble collagen peptide from Tilapia skin,ASCPT)的制备及抗菌性能。方法罗非鱼皮经浸提、离心、透析和冷冻干燥等步骤获得干燥的PSCPT、ASCPT。采用含有PSCPT或ASCPT的胰蛋白胨大豆肉汤培养基振荡培养大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌和白色念珠菌,定时测定细菌培养液的吸光度值、菌数、菌体细胞液电导率和细菌的形态特征。结果以牛胶原蛋白(Bovine skin collagen,BSC)为对照,PSCPT、ASCPT对大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌、白色念珠菌均有不同程度的生长抑制作用,抑制率从22.97%±2.35%到65.21%±6.42%,其中PSCPT抑制铜绿假单胞菌作用显著,抑制率达到65.21%±6.42%;ASCPT抑制大肠杆菌作用微弱,抑制率只有22.97%±2.35%。血球计数板法得到的抑菌效果类似吸光度值得到的结果,但是抑制率存在差别。PSCPT、ASCPT造成大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌、白色念珠菌的菌体细胞液电导率从36.42±1.35μs/cm变化到185.80±4.70μs/cm,其中PSCPT使铜绿假单胞菌的电导率低至36.42±1.35μs/cm,ASCPT使白色念珠菌的电导率达到185.80±4.70μs/cm。电镜下观察到抑菌率高和电导率低的菌株出现菌体变形和细胞壁破裂等现象。结论与BSC相比,PSCPT、ASCPT从细菌抑制率、菌体细胞液电导率和细菌的形态特征体现出一定的抑菌特性。
Objective To investigate the preparation and antibacterial activity of Pepsin soluble collagen peptide from Tilapia skin (PSCPT) and Acid soluble collagen peptide from Tilapia skin (ASCPT). Methods Tilapia skin was extracted, centrifuged, dialyzed and lyophilized to obtain dried PSCPT and ASCPT. Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans were cultured by shaking with Tryptone Soy Broth containing PSCPT or ASCPT, and the absorbance value, bacterial count, bacterial cell liquid conductivity Rate and bacterial morphological characteristics. Results Bovine skin collagen (BSC) was used as control. PSCPT and ASCPT all inhibited the growth of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans to various degrees with the inhibitory rate from 22.97% ± 2.35% to 65.21% ± 6.42%. Among them, the effect of PSCPT on Pseudomonas aeruginosa was significant, with the inhibition rate of 65.21% ± 6.42%. ASCPT inhibited Escherichia coli and the inhibitory rate was only 22.97% ± 2.35%. The antibacterial effect obtained by the blood counting plate method is similar to that obtained by the absorbance value, but there is a difference in the inhibition rate. PSCPT, ASCPT caused E. coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans bacterial cell liquid conductivity from 36.42 ± 1.35μs / cm to 185.80 ± 4.70μs / cm, which PSCPT so that the pseudo-green single The cell conductivity was as low as 36.42 ± 1.35 μs / cm, and the conductivity of Candida albicans was 185.80 ± 4.70 μs / cm with ASCPT. Under the electron microscope, the strain with high antibacterial activity and low conductivity showed cell deformation and cell wall rupture. Conclusion Compared with BSC, PSCPT and ASCPT showed some antibacterial properties from the inhibition rates of bacteria, bacterial cell liquid conductivity and the morphological characteristics of bacteria.