Kim W K, Patterson P H
Department of Poultry Science, The Pennsylvania State University, University Park, Pennsylvania 16802-3501, USA.
Poult Sci. 2003 Oct;82(10):1554-8. doi: 10.1093/ps/82.10.1554.
Ammonia gas produced from poultry manure can be a potential source of environmental pollution. Microbial uricase in poultry manure is an important target enzyme to reduce ammonia production because ammonia is mainly generated from the microbial decomposition of uric acid in the manure. Thus, the inhibition of microbial uricase is critical in preventing NH3 volatilization. A potential method of inhibiting uricase activity is the use of antibodies specific to microbial uricase. Therefore, this study was designed to evaluate 1) the production of the uricase-specific egg yolk antibody (IgY) from immunized hens and 2) the effect of the uricase-specific IgY on the activity of uricase. A total of 12 Single Comb White Leghorn hens were injected intramuscularly with uricase from Arthrobacter globiformis. The hens were immunized a second and third time at 1 and 2 wk, respectively, after the initial injection. The production of uricase-specific IgY was first detected at 2 wk after the initial immunization, and levels increased more than threefold at 4 wk. The method including water extraction, ammonium sulfate precipitation, and ethanol precipitation showed the most acceptable IgY purity and over 97% uricase-specific IgY recovery. Finally, the effect of the uricase-specific IgY on uricase activity was determined by a uricase assay. The slope ratio showed that the immune-IgY from the uricase-immunized hens and nonimmune IgY from nonimmunized control hens reduced uricase degradation by 58 and 43%, respectively, compared to the uricase treatment. The regression slope indicated that the immune IgY and nonimmune IgY treatments had a significant inhibitory effects on uricase activity compared to the uricase.
家禽粪便产生的氨气可能是环境污染的一个潜在来源。家禽粪便中的微生物尿酸酶是减少氨气产生的重要靶标酶，因为氨气主要是由粪便中尿酸的微生物分解产生的。因此，抑制微生物尿酸酶对于防止氨气挥发至关重要。抑制尿酸酶活性的一种潜在方法是使用针对微生物尿酸酶的抗体。因此，本研究旨在评估：1）免疫母鸡产生的尿酸酶特异性蛋黄抗体（IgY）；2）尿酸酶特异性IgY对尿酸酶活性的影响。总共12只单冠白来航母鸡通过肌肉注射球形节杆菌的尿酸酶进行免疫。在初次注射后第1周和第2周，母鸡分别进行第二次和第三次免疫。初次免疫后2周首次检测到尿酸酶特异性IgY的产生，4周时水平增加了三倍多。包括水提取、硫酸铵沉淀和乙醇沉淀的方法显示出最可接受的IgY纯度和超过97%的尿酸酶特异性IgY回收率。最后，通过尿酸酶测定法确定尿酸酶特异性IgY对尿酸酶活性的影响。斜率比表明，与尿酸酶处理相比，尿酸酶免疫母鸡的免疫IgY和未免疫对照母鸡的非免疫IgY分别使尿酸酶降解降低了58%和43%。回归斜率表明，与尿酸酶相比，免疫IgY和非免疫IgY处理对尿酸酶活性具有显著的抑制作用。
