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孵化场蛋壳膜的蛋白质谱

Protein profiles of hatchery egg shell membrane.

作者信息

Rath N C, Liyanage R, Makkar S K, Lay J O

机构信息

USDA/Agricultural Research Service, Poultry Production and Product Safety Research Unit, Fayetteville, AR 72701 USA.

Statewide Mass Spectrometry Facility, University of Arkansas, Fayetteville, AR 72701 USA.

出版信息

Proteome Sci. 2017 Mar 3;15:4. doi: 10.1186/s12953-017-0112-6. eCollection 2016.

Abstract

BACKGROUND

Eggshells which consist largely of calcareous outer shell and shell membranes, constitute a significant part of poultry hatchery waste. The shell membranes (ESM) not only contain proteins that originate from egg whites but also from the developing embryos and different contaminants of microbial and environmental origins. As feed supplements, during post hatch growth, the hatchery egg shell membranes (HESM) have shown potential for imparting resistance of chickens to endotoxin stress and exert positive health effects. Considering that these effects are mediated by the bioactive proteins and peptides present in the membrane, the objective of the study was to identify the protein profiles of hatchery eggshell membranes (HESM).

METHODS

Hatchery egg shell membranes were extracted with acidified methanol and a guanidine hydrochloride buffer then subjected to reduction/alkylation, and trypsin digestion. The methanol extract was additionally analyzed by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). The tryptic digests were analyzed by liquid chromatography and tandem mass spectrometry (LC-MS-MS) to identify the proteins.

RESULTS

Our results showed the presence of several proteins that are inherent and abundant in egg white such as, ovalbumin, ovotransferrin, ovocleidin-116, and lysozyme, and several proteins associated with cytoskeletal, cell signaling, antimicrobial, and catalytic functions involving carbohydrate, nucleic acid, and protein metabolisms. There were some blood derived proteins most likely originating from the embryos and several other proteins identified with different aerobic, anaerobic, gram positive, gram negative, soil, and marine bacterial species some commensals and others zoonotic.

CONCLUSION

The variety of bioactive proteins, particularly the cell signaling and enzymatic proteins along with the diverse microbial proteins, make the HESM suitable for nutritional and biological application to improve post hatch immunity of poultry.

摘要

背景

蛋壳主要由钙质外壳和壳膜组成,是家禽孵化场废弃物的重要组成部分。壳膜(ESM)不仅含有源自蛋清的蛋白质,还含有来自发育中胚胎的蛋白质以及微生物和环境来源的不同污染物。作为饲料补充剂,在孵化后生长期间,孵化场蛋壳膜(HESM)已显示出赋予鸡对内毒素应激的抵抗力并产生积极健康影响的潜力。鉴于这些影响是由膜中存在的生物活性蛋白质和肽介导的,本研究的目的是鉴定孵化场蛋壳膜(HESM)的蛋白质谱。

方法

用酸化甲醇和盐酸胍缓冲液提取孵化场蛋壳膜,然后进行还原/烷基化和胰蛋白酶消化。甲醇提取物另外通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)进行分析。通过液相色谱和串联质谱(LC-MS-MS)分析胰蛋白酶消化产物以鉴定蛋白质。

结果

我们的结果显示存在几种蛋清中固有的丰富蛋白质,如卵清蛋白、卵转铁蛋白、卵壳蛋白-116和溶菌酶,以及几种与细胞骨架、细胞信号传导、抗菌和涉及碳水化合物、核酸和蛋白质代谢的催化功能相关的蛋白质。有一些可能源自胚胎的血液衍生蛋白质,以及其他几种与不同的需氧、厌氧、革兰氏阳性菌(革兰氏阴性菌)、土壤和海洋细菌物种鉴定出的蛋白质,其中一些是共生菌,其他是人畜共患病菌。

结论

多种生物活性蛋白质,特别是细胞信号传导和酶蛋白以及多样的微生物蛋白质,使HESM适用于营养和生物学应用,以提高家禽孵化后的免疫力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f334/5335749/38dffc955dc8/12953_2017_112_Fig1_HTML.jpg

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