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采用三氯乙酸裂解法分离双歧杆菌细胞壁肽聚糖,得到的肽聚糖约占双歧杆菌细胞干重的8.4%,其中总糖含量约为24.2%,脂肪酸含量约为3.2%,蛋白质含量约为2.4%;氨基酸含量及组分分析结果显示:与N乙酰胞壁酸羧基连接的肽链上的甘氨酸、谷氨酸、赖氨酸和丙氨酸间的摩尔比为1∶0.7∶1.1∶6.7。将所得肽聚糖经超声波破碎后,制成0.1%和1%两种浓度的悬液,并将相应的缓冲液设置为对照组,分别注射给日本对虾和牙鲆,测定其对日本对虾和牙鲆血清酚氧化酶(PO)、超氧化物歧化酶(SOD)、碱性磷酸酶(ALP)、酸性磷酸酶(ACP)和溶菌酶(LSZ)5种免疫酶活性的影响。结果显示:肽聚糖可显著提高日本对虾血清内的PO活性和LSZ活性,并在96h内一直维持在较高水平;肽聚糖可适当提高日本对虾血清SOD活性,但到72h又降回原来的水平;肽聚糖可影响日本对虾血清的ALP活性和ACP活性,且两种作用趋势相近,96h内呈逐渐平缓上升的趋势。肽聚糖可显著提高牙鲆血清的LSZ活性,但这种升高具有明显的时效性,72h后降至原来的水平,且不同浓度的作用效果也有差异。由此可见,肽聚糖可作为免疫增强剂用于提高养殖日本对虾和牙鲆的非特异性免疫水平。
The trichloracetic acid cleavage method was used to separate bifidobacteria cell wall peptidoglycan. The obtained peptidoglycan accounted for about 8.4% of the dry weight of Bifidobacterium cells, of which the total sugar content was about 24.2% and the fatty acid content was about 3.2%. The protein content About 2.4%; amino acid content and composition analysis showed that: the N-acetyl-linked muramic acid carboxyl linked peptide chain glycine, glutamic acid, lysine and alanine molar ratio of 1: 0.7: 1.1 : 6.7. After the resulting peptidoglycan was disrupted by ultrasonic wave, two concentrations of 0.1% and 1% suspension were prepared, and the corresponding buffer was set as the control group, which were respectively injected to Japanese shrimp and Japanese flounder, Effects of 5 kinds of immune enzymes activities of serum floss oxidase (PO), superoxide dismutase (SOD), alkaline phosphatase (ALP), acid phosphatase (ACP) and lysozyme (LSZ) The results showed that peptidoglycan significantly increased the activity of PO and LSZ in serum of Penaeus vannamei and maintained at a high level within 96 hours. Peptidoglycan could increase SOD activity in serum of Penaeus japonicus, but returned to the original value at 72 hours Peptidoglycan could affect ALP activity and ACP activity of Japanese shrimp serum, and the two effects tended to be similar and gradually increased gradually within 96 hours. Peptidoglycan significantly increased the LSZ activity of flounder serum, but this increase has obvious timeliness, after 72h to the original level, and the effect of different concentrations are also different. Thus, peptidoglycans can be used as an immune enhancer to improve the non-specific immunity of Japanese shrimp and Japanese flounder.