不同畜禽骨蛋白肽的制备、理化特性表征及其生物活性

doi:10.3864/j.issn.0578-1752.2022.13.012

摘要/Abstract

摘要：

【目的】制备骨蛋白肽（livestock and poultry bone peptides,LBPs）用于功能性骨源食品的开发是畜禽骨副产物高值化加工利用的重要途径之一。选取4种主要畜禽骨原料基于相同工艺分别制备LBPs,对比分析其理化特性与生物活性,为功能性骨源食品的开发及畜禽骨资源的高值化加工利用提供参考。【方法】以牦牛腿骨、黄牛腿骨、猪腿骨及鸡腿骨为原料分别制备牦牛骨蛋白肽（yak bone peptides,YBPs）、黄牛骨蛋白肽（bovine bone peptides,BBPs）、猪骨蛋白肽（porcine bone peptides,PBPs）、鸡骨蛋白肽（chicken bone peptides,CBPs）,并对4种LBPs基本营养组分、氨基酸组成、分子量分布、粒径分布等理化性质进行表征;对比分析4种LBPs的促成骨细胞增殖、促巨噬细胞增殖、血管紧张素转换酶抑制（angiotensin converting enzyme inhibitors,ACEI）、抗氧化等生物活性差异。【结果】YBPs、BBPs和PBPs粗蛋白含量分别为（89.70±0.77）%、（90.43±0.88）%和（89.36±1.32）%,显著高于CBPs（(79.18±1.49)%）;4种LBPs均缺乏色氨酸,CBPs的必需氨基酸和含硫氨基酸均显著高于YBPs、BBPs和PBPs;4种LBPs主要由MW<2 kD的小分子多肽片段组成,占比约90%;LBPs粉体粒径分布差异不显著,主要集中于10—20 µm和40—60 µm范围内;不同种源LBPs生物活性特征分析发现：YBPs促成骨细胞增殖效应最显著,0.5 mg∙mL^-1时的促增殖率达37.27%;BBPs促巨噬细胞增殖效应最强,5 mg∙mL^-1时的促增殖率达39.26%;PBPs的ACEI活性最强,15 mg∙mL^-1时ACE活性抑制率为82.37%;与BBPs、PBPs和CBPs相比,YBPs的综合抗氧化能力最强。【结论】4种不同种源LBPs的理化特性存在一定的差异,但符合功能性骨源食品的原料要求。不同种源LBPs的生物活性存在一定的差异,因而适宜于不同功能性骨源食品的开发：YBPs的促成骨细胞增殖效应及综合抗氧化能力最强,更适宜于改善骨健康及抗氧化类功能性骨源食品的开发;BBPs的促巨噬细胞增殖效应最强,更适宜于开发免疫调节类功能性骨源食品;PBPs的ACEI活性最高,更适宜于开发血压控制类功能性骨源食品;CBPs具备更优良的粉体性质及更高的矿物质含量,可用作膳食营养补充剂。

关键词: 畜禽骨, 骨蛋白肽, 理化特性, 生物活性, 高值化利用, 功能性骨源食品

Abstract:

【Objective】The preparation of bone peptides (livestock and poultry bone peptides, LBPs) for the development of bone-derived functional foods is one of the important ways for the high-value utilization of livestock and poultry bone by-products. In this experiment, the leg bone from four main livestock and poultry species were used as raw materials to prepare LBPs based on the same process, and their physicochemical properties and bioactivities were compared and analyzed, so as to provide the reference for the high-value utilization of livestock and poultry bone resources and the development of bone-derived functional foods.【Method】 Yak bone peptides (YBPs), bovine bone peptides (BBPs), porcine bone peptides (PBPs) and chicken bone peptides (CBPs) were prepared from yak, bovine, porcine and chicken leg bones, respectively. Physicochemical properties were characterized, including basic nutritional components, amino acid compositions, molecular weight and particle size distribution. Moreover, the bioactivities of the four LBPs, including osteoblast promoting proliferation, immune regulation, angiotensin converting enzyme inhibitors (ACEI) and antioxidation, were compared and analyzed. 【Result】 The relative contents of crude protein in YBPs, BBPs and PBPs were (89.70±0.77)%, (90.43±0.88)% and (89.36±1.32)%, respectively, which were significant higher than that of CBPs (79.18±1.49)%). The essential amino acids and sulfur-amino acids of CBPs were significant higher than those of YBPs, BBPs and PBPs. The four LBPs were mainly composed of small molecular peptides with MW<2 kD, accounting for about 90%. There was no significant difference in particle size distribution of LBPs powder, which was mainly concentrated in the range of 10-20 µm and 40-60 µm. The bioactivities of different LBPs were analyzed. It was found that YBPs had the most significant osteoblasts promoting proliferation effect, and the proliferation rate was 37.27% at 0.5 mg∙mL^-1; BBPs had the strongest effect on macrophage proliferation, and the proliferation rate was 39.26% at 5 mg∙mL^-1; PBPs had the strongest ACEI activity, and the inhibition rate of ACE activity was 82.37% at 15 mg∙mL^-1; YBPs had the strongest comprehensive antioxidant capacity, compared with BBPs, PBPs and CBPs. 【Conclusion】The physicochemical properties of LBPs from four distinct species were different, but they all met the demand of raw material for the development of bone-derived functional foods. The bioactivities of LBPs from different species were different, which were suitable for the development of different bone-derived functional foods: YBPs had the strongest osteoblasts promoting proliferation effect and comprehensive antioxidant capacity, which was more suitable for the development of bone health improving or antioxidant bone-derived functional foods; BBPs had the strongest effect on promoting macrophage proliferation, which was more suitable for the development of immunomodulatory bone-derived functional foods; PBPs had the strongest ACEI activity and was more suitable for the development of bone-derived functional foods with blood pressure control effect; CBPs had the better powder properties and higher mineral content, which could be used as dietary nutritional supplements.

Key words: livestock and poultry bone, bone peptides, physicochemical properties, bioactivities, high value utilization, bone- derived functional foods

刘泓,郭玉杰,许雄,李侠,张鸿儒,齐立伟,孙雪梅,张春晖. 不同畜禽骨蛋白肽的制备、理化特性表征及其生物活性[J]. 中国农业科学, 2022, 55(13): 2629-2642.

LIU Hong,GUO YuJie,XU Xiong,LI Xia,ZHANG HongRu,QI LiWei,SUN XueMei,ZHANG ChunHui. Preparation, Physicochemical Characterization and Bioactivity Comparison of Different Livestock and Poultry Bone Peptides[J]. Scientia Agricultura Sinica, 2022, 55(13): 2629-2642.

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https://www.chinaagrisci.com/CN/Y2022/V55/I13/2629

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畜禽骨蛋白肽的氨基酸组成"

氨基酸 Amino acids	FAO/WHO推荐摄入量 FAO/WHO recommended intake (mg/kg body weight/day)	含量 Content (g/100 g)
氨基酸 Amino acids		YBPs	BBPs	PBPs	CBPs
天冬酰胺Asp	-	5.08±0.27a	5.21±0.40a	5.07±0.27a	5.27±0.09a
谷氨酸Glu	-	9.75±0.35ab	9.29±0.75b	9.39±0.41b	10.47±0.21a
苏氨酸Thr*	7	1.82±0.05b	1.69±0.15b	1.79±0.09b	2.20±0.04a
丝氨酸Ser	-	2.80±0.12a	2.69±0.24a	2.70±0.06a	2.24±0.05b
甘氨酸Gly	-	19.6±0.61b	22.64±1.13a	20.05±1.30b	14.26±0.20c
丙氨酸Ala	-	8.26±0.24b	9.18±0.21a	8.13±0.39b	6.65±0.10c
脯氨酸Pro	-	10.4±1.03a	11.07±0.64a	10.95±0.53a	8.03±0.16b
羟脯氨酸Hyp	-	9.39±0.85a	9.91±1.58a	8.37±00.91b	7.46±0.33c
苯丙氨酸Phe*	14	1.85±0.08a	1.78±0.08a	1.93±0.12a	1.91±0.05a
酪氨酸Tyr	14	0.68±0.01a	0.54±0.02b	0.63±0.05a	0.57±0.02b
半胱氨酸Cys	13	0.02±0.01a	0.02±0.00a	0.05±0.01a	0.05±0.00a
蛋氨酸Met*	13	0.75±0.03b	0.74±0.02b	0.73±0.01b	1.04±0.07a
缬氨酸Val*	10	2.44±0.08a	2.49±0.07a	2.47±0.17a	2.04±0.04b
异亮氨酸Ile*	10	1.46±0.05b	1.43±0.10b	1.21±0.08c	1.83±0.07a
亮氨酸Leu*	14	3.34±0.11a	3.05±0.19b	3.00±0.18b	3.61±0.11a
赖氨酸Lys*	12	3.29±0.10a	3.08±0.21a	3.22±0.20a	3.39±0.06a
羟赖氨酸Hyl	-	1.13±0.11a	1.09±0.06a	0.69±0.03b	0.56±0.02c
组氨酸His	-	0.84±0.03a	0.69±0.01b	0.75±0.03b	0.87±0.04a
精氨酸Arg	-	6.81±0.18b	7.32±0.22a	6.62±0.41b	5.44±0.13c
总计 Total
氨基酸总量 Total amino acids	-	89.81	93.91	87.75	77.32
含硫氨基酸¹⁾Sulphur-amino acids		0.77	0.76	0.78	1.09
芳香族氨基酸²⁾ Aromatic amino acids		2.53	2.32	2.56	2.48
疏水性氨基酸³⁾Hydrophilic amino acids		28.59	29.74	28.42	24.54
亲水性氨基酸⁴⁾ Hydrophobic amino acids		50.70	53.17	50.27	44.76
亚氨基酸⁵⁾ Imino acid		19.88	20.98	19.32	15.49
必需氨基酸* Essential amino acids		17.40	16.65	16.82	18.54
氨基酸比值系数分 SRC	100	72.81	73.44	71.17	53.67

表3

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样品 Sample	水分 Moisture (%)	矿物质 Minerals (%)	粗蛋白 Crude protein (%)	粗脂肪Crude fat (%)
YBPs	4.96±0.24b	4.18±0.18b	89.70±0.77a	0.17±0.01a
BBPs	4.46±0.61b	4.30±0.51b	90.43±0.88a	0.16±0.01a
PBPs	4.80±0.49b	4.68±0.45b	89.36±1.32a	0.13±0.02b
CBPs	5.77±0.52a	12.9±0.80a	79.18±1.49b	0.07±0.01c