In this paper, bovine bone collagen peptides were extracted by ultrasound-assisted enzymatic hydrolysis and their antioxidant properties were investigated. Bovine bone was used as the raw material, the hydrolysis degree of proteins was used as the evaluation index, and the optimal extraction process conditions were optimized on the basis of a single factor combined with response surface methodology. The composition of free amino acids and molecular mass distribution of peptides in the peptide solution were investigated, and the scavenging abilities of DPPH radicals and superoxide anion radicals were measured at a concentration gradient of 1-5 mg/mL in the peptide solution to evaluate the antioxidant activity of bovine bone collagen peptide. Results showed that under the enzymatic hydrolysis conditions of ultrasonic power of 400 W, ultrasonic time of 120 min, substrate concentration of 4.4 g/100 mL, and enzyme dosage of 6 000 U/g, the degree of hydrolysis of bovine bone collagen was 20.94%, which was close to the predicted value of 21.33%. The peptide solution was rich in hydrophobic and essential amino acids, with 86.83% of peptides below 1 000 Da. The results of antioxidant indexes in vitro showed that the highest scavenging rate of DPPH radicals was 43.80%, and the highest scavenging rate of superoxide anion radicals was 64.74%, both of which were significantly higher than that of the non-sonication group at 37.33% and 57.41% (P<0.05), indicating that the peptide solution extracted by ultrasonic-assisted enzymatic hydrolysis had significant antioxidant properties. Therefore, extraction of bovine bone collagen peptides using ultrasonic-assisted enzymatic hydrolysis is a green and efficient extraction method, which is of great significance for the utilization of bovine bone proteins and the development of active peptides.
[1] SONG S Q, LI S S, FAN L, et al.A novel method for beef bone protein extraction by lipase-pretreatment and its application in the Maillard reaction[J].Food Chemistry, 2016, 208:81-88.
[2] CHIANG J H, HARDACRE A K, PARKER M E.Effects of Maillard-reacted beef bone hydrolysate on the physicochemical properties of extruded meat alternatives[J].Journal of Food Science, 2020, 85(3):567-575.
[3] 刘斯汝, 罗龙龙, 任卫合, 等.牛肉酶解工艺优化及其氨基酸组成分析[J].肉类研究, 2020, 34(11):21-26.
LIU S R, LUO L L, REN W H, et al.Optimization of enzymatic hydrolysis of beef and analysis of amino acid composition of hydrolysate[J].Meat Research, 2020, 34(11):21-26.
[4] SHEN D Y, BEGUM N, SONG H L, et al.In vitro and in vivo antioxidant activity and umami taste of peptides (<1 kDa) from porcine bone protein extract[J].Food Bioscience, 2021, 40:100901.
[5] 李晓瑞, 王梓, 刘贵珊, 等.组合酶对复合骨素酶解液呈味物质的影响[J].食品工业科技, 2020, 41(22):50-56.
LI X R, WANG Z, LIU G S, et al.Effects of combined enzymes on flavoring substances of compound osteolysin hydrolysate[J].Science and Technology of Food Industry, 2020, 41(22):50-56.
[6] 李露园, 王升帆, 朱有贵, 等.酶法制备鲟鱼皮胶原蛋白多肽及其抗氧化活性研究[J].食品与发酵工业, 2019, 45(20):138-143.
LI L Y, WANG S F, ZHU Y G, et al.Enzymatic preparation and antioxidant activity of sturgeon skin collagen polypeptide[J].Food and Fermentation industries, 2019, 45(20):138-143.
[7] CHI C F, WANG B, HU F Y, et al.Purification and identification of three novel antioxidant peptides from protein hydrolysate of bluefin leatherjacket (Navodon septentrionalis) skin[J].Food Research International, 2015, 73:124-129.
[8] 刘倩霞, 刘东, 张俊, 等.2种蛋白酶酶解曲拉干酪素条件优化及抗氧化性比较[J].食品科学, 2019, 40(8):225-234;218.
LIU Q X, LIU D, ZHANG J, et al.Optimization of hydrolysis conditions of Qula casein with two proteases and comparative antioxidant activity of hydrolysates[J].Food Science, 2019, 40(8):225-234;218.
[9] 魏洁琼, 余群力, 韩玲, 等.牛骨胶原蛋白肽制备工艺优化及抗氧化活性分析[J].甘肃农业大学学报, 2020, 55(5):203-211;218.
WEI J Q, YU Q L, HAN L, et al.Optimization of preparation process and antioxidant activity analysis of bovine bone collagen peptide[J].Journal of Gansu Agricultural University, 2020, 55(5):203-211;218.
[10] UMEGO E C, HE R H, REN W B, et al.Ultrasonic-assisted enzymolysis:Principle and applications[J].Process Biochemistry, 2021, 100:59-68.
[11] 刘伟. 超声波处理对河蚌肉酶解物美拉德反应的影响及其微囊化调味料制备[D].长春:吉林大学, 2019.
LIU W.Effect of ultrasonic treatment on Maillard reaction of mussel meat hydrolysate and preparation of microencapsulated seasoning[D].Changchun:Jilin University, 2019.
[12] DONG Z Y, LI M Y, TIAN G, et al.Effects of ultrasonic pretreatment on the structure and functionality of chicken bone protein prepared by enzymatic method[J].Food Chemistry, 2019, 299:125103.
[13] 董海英. 鸭骨的酶法水解及产物的初步应用研究[D].武汉:武汉工业学院, 2010.
DONG H Y.Enzymatic hydrolysis of duck bone and prelimlinary application of the product[D].Wuhan:Wuhan Polytechnic University, 2010.
[14] YANG L H, GUO Z L, WEI J Q, et al.Extraction of low molecular weight peptides from bovine bone using ultrasound-assisted double enzyme hydrolysis:Impact on the antioxidant activities of the extracted peptides[J].LWT, 2021, 146:111470.
[15] 李云嵌, 杨曦, 刘江, 等.超声波辅助碱法提取美藤果分离蛋白及其加工性质研究[J].食品与发酵工业, 2021, 47(9):128-135.
LI Y Q, YANG X, LIU J, et al.Ultrasonic-assisted alkaline extraction of Plukenetia volubilis protein isolate and its processing properties[J].Food and Fermentation Industry, 2021, 47(9):128-135.
[16] YAMAMOTO S, DEGUCHI K, ONUMA M, et al.Absorption and urinary excretion of peptides after collagen tripeptide ingestion in humans[J].Biological & Pharmaceutical Bulletin, 2016, 39(3):428-434.
[17] 王子涵, 向敏, 徐茂, 等.响应面优化黑果腺肋花楸汁澄清工艺及其抗氧化活性评价[J].食品与发酵工业, 2021, 47(8):189-196.
WANG Z H, XIANG M, XU M, et al.Optimization of clarification process of Aornia melanocarpa juice by response surface methodology and its antioxidant activity evaluation[J].Food and Fermentation Industries, 2021, 47(8):189-196.
[18] 南学敏. 羊骨胶原蛋白肽抗氧化活性及氨基酸组成分析[D].呼和浩特:内蒙古农业大学, 2019.
NAN X M.Analysis of antioxidant activity and amino acid composition of sheep bone collagen peptide[D].Hohhot:Inner Mongolia Agricultural University, 2019.
[19] YU L N, SUN J, LIU S F, et al.Ultrasonic-assisted enzymolysis to improve the antioxidant activities of peanut (Arachin conarachin L.) antioxidant hydrolysate[J].International Journal of Molecular Sciences, 2012, 13(7):9 051-9 068.
[20] RAGHAVAN S,KRISTINSSON H G. ACE-inhibitory activity of tilapia protein hydrolysates[J].Food Chemistry, 2009, 117(4):582-588.
