Ultrasound-assisted enzymolytic extraction of chlorohemin from yak blood powder: response surface optimization and quality characterization

doi:10.3969/j.issn.1004-1524.20230926

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (6): 1357-1367.DOI: 10.3969/j.issn.1004-1524.20230926

• Food Science • Previous Articles Next Articles

Ultrasound-assisted enzymolytic extraction of chlorohemin from yak blood powder: response surface optimization and quality characterization

ZHANG Jin¹(), WU Xiaoli¹^,², TIAN Yuwei³, ZHAO Ke¹, LI Huanhuan¹, Dase ⁴, Cirendajie ⁴, CHEN Lihong¹, TANG Honggang¹^,^*()

1. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Food Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. College of Food and Pharmacy, Zhejiang Ocean University, Zhoushan 316022, Zhejiang, China
3. School of Science, Xingzhi College Zhejiang Normal University, Jinhua 310018, Zhejiang, China
4. Naqu Gonyal Animal Husbandry Technology and Industry Company, Naqu 852014, Xizang, China

Received:2023-07-28 Online:2024-06-25 Published:2024-07-02

Abstract

Abstract:

The objective of this work was to extract, optimize, and evaluate the chlorohemin from yak blood powder using ultrasonic-assisted enzymolytic technology. The ultrasound-assisted enzymolytic time, enzyme/substrate concentration, and ultrasound specific power were the independent variables, and the purity was the response variable. The Box-Behnken response surface design was also used to optimize the extraction process. Furthermore, the purity, yield, color, and chemical structure of the extracted chlorohemin with response surface optimization were characterized by ultraviolet-visible spectrophotometer, colorimeter, and Fourier transform-infrared spectrometer, respectively. The results showed that ultrasound-assisted enzymolytic time, enzyme/substrate concentration, and ultrasound specific power had significant effects on the purity of chlorohemin extract (P<0.05). The optimal extraction conditions were determined as ultrasound-assisted enzymatic time 4 h, enzyme/substrate concentration 17 U·mg^-1, and ultrasound specific power 800 W·L^-1 after response surface optimization. The purity of extracted chlorohemin under these conditions was 14.89%, the extraction yield was 63.30%. Compared with the chlorohemin extracted through normal enzymolysis (without ultrasound-assisted treatment) under the same conditions, the yield and purity were increased by 1.29 and 1.07 times (P<0.05), respectively. Moreover, there was not significant difference in chemical structure between these two samples. However, the color of extracted chlorohemin with ultrasound-assisted enzymolysis and response surface optimization was closer to the standard substance of chlorohemin. The ultrasound-assisted treatment (4 h at 800 W·L^-1 ultrasound specific power) could significantly improve the purity and yield of chlorohemin extraction from yak blood powder without changing its physicochemical properties.

Key words: yak blood, ultrasound-assisted enzymolysis, chlorohemin, response surface optimization, Fourier transform-infrared spectroscopy

CLC Number:

TS251.93

ZHANG Jin, WU Xiaoli, TIAN Yuwei, ZHAO Ke, LI Huanhuan, Dase , Cirendajie , CHEN Lihong, TANG Honggang. Ultrasound-assisted enzymolytic extraction of chlorohemin from yak blood powder: response surface optimization and quality characterization[J]. Acta Agriculturae Zhejiangensis, 2024, 36(6): 1357-1367.

Figures/Tables 11

References 31

[1]	杨柏高, 郝海生, 杜卫华, 等. 牦牛高原适应研究进展[J]. 畜牧兽医学报, 2023, 54(1): 12-23.
	YANG B G, HAO H S, DU W H, et al. Advances in research on plateau adaptation of yak[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 12-23. (in Chinese with English abstract)
[2]	闫忠心, 靳义超, 李升升. 青藏高原牧区风干牦牛肉的营养品质分析[J]. 青海畜牧兽医杂志, 2017, 47(1): 20-23, 8.
	YAN Z X, JIN Y C LI S S. Analysis of nutritional quality of dry yak meat in Qinghai-Tibet Plateau[J]. Chinese Qinghai Journal of Animal and Veterinary Sciences, 2017, 47(1): 20-23, 8. (in Chinese with English abstract)
[3]	DIAO N C, GONG Q L, LI J M, et al. Prevalence of bovine viral diarrhea virus (BVDV) in yaks between 1987 and 2019 in mainland China: a systematic review and meta-analysis[J]. Microbial Pathogenesis, 2020, 144: 104185.
[4]	ZHANG J, LI X Q, ZHAO K, et al. In vitro digestion and fermentation combined with microbiomics and metabolomics reveal the mechanism of superfine yak bone powder regulating lipid metabolism by altering human gut microbiota[J]. Food Chemistry, 2023, 410: 135441.
[5]	HUANG Q, DONG K, WANG Q, et al. Changes in volatile flavor of yak meat during oxidation based on multi-omics[J]. Food Chemistry, 2022, 371: 131103.
[6]	MEDHI D, SANTRA A, PAUL V, et al. Effect of plane of nutrition on blood biochemical parameters and attainment of sexual maturity in growing yaks[J]. The Indian Journal of Animal Sciences, 2018, 88(4): 470-473.
[7]	高子涵, 张卫兵, 梁琪, 等. 牦牛血粉制备氯化血红素工艺优化研究[J]. 食品工业科技, 2013, 34(19): 248-251.
	GAO Z H, ZHANG W B, LIANG Q, et al. Optimization of technology for hemin from Bosgruniens blood powder[J]. Science and Technology of Food Industry, 2013, 34(19): 248-251. (in Chinese with English abstract)
[8]	王丽媛, 高艳蕾, 张丽, 等. 畜禽副产物的加工利用现状及研究展望[J]. 食品科技, 2022, 47(6): 174-183.
	WANG L Y, GAO Y L, ZHANG L, et al. Processing and utilization status and research prospect of livestock and poultry by-products[J]. Food Science and Technology, 2022, 47(6): 174-183. (in Chinese with English abstract)
[9]	胡睿琦, 王建敏, 王楠, 等. 冰醋酸法提取猪血红素的研究[J]. 安徽农业科学, 2014, 42(16): 5211-5213.
	HU R Q, WANG J M, WANG N, et al. Study on the extraction of hemoglobin from porcine blood by acetic acid method[J]. Journal of Anhui Agricultural Sciences, 2014, 42(16): 5211-5213. (in Chinese with English abstract)
[10]	沈亮, 徐晓燕, 赵斌斌, 等. 改良酸性丙酮法提取鸭血血红素的研究[J]. 中国药学杂志, 2014, 49(8): 664-668.
	SHEN L, XU X Y, ZHAO B B, et al. Preparation of heme from duck blood by a new acidified acetone extraction method[J]. Chinese Pharmaceutical Journal, 2014, 49(8): 664-668. (in Chinese with English abstract)
[11]	代文婷, 陶永霞, 杨海燕, 等. 超声波结合CMC-Na对马血中血红素提取工艺效果影响[J]. 食品工业科技, 2014, 35(13): 257-261.
	DAI W T, TAO Y X, YANG H Y, et al. The impact of ultrasound combined with CMC-Na on the extraction of heme from horse blood[J]. Science and Technology of Food Industry, 2014, 35(13): 257-261. (in Chinese with English abstract)
[12]	贾志春, 张珍, 张盛贵, 等. 木瓜蛋白酶酶解牦牛血红蛋白制备氯化血红素关键工艺研究[J]. 食品工业科技, 2016, 37(3): 206-210, 215.
	JIA Z C, ZHANG Z, ZHANG S G, et al. Study on the preparation technology of hemin from enzymolysis yak blood by papain[J]. Science and Technology of Food Industry, 2016, 37(3): 206-210, 215. (in Chinese with English abstract)
[13]	韩晴, 王维婷, 王守经, 等. 羊血亚铁血红素的酶法制备及抗氧化活性研究[J]. 食品科技, 2023, 48(3): 147-154.
	HAN Q, WANG W T, WANG S J, et al. Enzymatic preparation and antioxidant activity of ferroheme from sheep blood[J]. Food Science and Technology, 2023, 48(3): 147-154. (in Chinese with English abstract)
[14]	TAVANANDI H A, RAGHAVARAO K S M S. Ultrasound-assisted enzymatic extraction of natural food colorant C-Phycocyanin from dry biomass of Arthrospira platensis[J]. LWT, 2020, 118: 108802.
[15]	于娇, 陈胜军, 胡晓, 等. 超声波辅助提取坛紫菜蛋白条件优化及其基础特性研究[J]. 食品与发酵工业, 2019, 45(8): 142-148.
	YU J, CHEN S J, HU X, et al. Extraction and characterization of protein from Porphyra haitanensis by optimized ultrasound-assisted extraction[J]. Food and Fermentation Industries, 2019, 45(8): 142-148. (in Chinese with English abstract)
[16]	闫忠心. 超声辅助提取冷冻牦牛血中血红素的工艺优化[J]. 青海大学学报, 2018, 36(5): 1-6.
	YAN Z X. Technology optimization of ultrasonic-assisted heme extraction from frozen yak blood[J]. Journal of Qinghai University, 2018, 36(5): 1-6. (in Chinese with English abstract)
[17]	张晋, 谭芦兰, 陈伊凡, 等. 可溶性膳食纤维强化重组蛋片的响应面优化制备和品质评价[J]. 中国食品学报, 2020, 20(12): 155-166.
	ZHANG J, TAN L L, CHEN Y F, et al. Response surface optimization and quality evaluation of soluble dietary fiber-fortified restructuring egg white curd[J]. Journal of Chinese Institute of Food Science and Technology, 2020, 20(12): 155-166. (in Chinese with English abstract)
[18]	ZIMET P, MOMBRÚ Á W, FACCIO R, et al. Optimization and characterization of nisin-loaded alginate-chitosan nanoparticles with antimicrobial activity in lean beef[J]. LWT, 2018, 91: 107-116.
[19]	YING Y B, XIANG Y C, LIU J L, et al. Optimization of ultrasonic-assisted freezing of Penaeus chinensis by response surface methodology[J]. Food Safety and Quality, 2021, 5(2):143-151.
[20]	ZHANG J, LI H H, CHEN Y F, et al. Microencapsulation of immunoglobulin Y: optimization with response surface morphology and controlled release during simulated gastrointestinal digestion[J]. Journal of Zhejiang University Science B, 2020, 21(8): 611-627.
[21]	MA Y Q, SHAN A S, WANG R H, et al. Characterization of egg white powder gel structure and its relationship with gel properties influenced by pretreatment with dry heat[J]. Food Hydrocolloids, 2021, 110: 106149.
[22]	刘振荣, 王君, 张向东, 等. 超声波法提取氯化血红素的初步研究[J]. 辽宁大学学报(自然科学版), 2003, 30(4): 373-377.
	LIU Z R, WANG J, ZHANG X D, et al. Primary investigation on the extraction of hemin using an ultrasonic method[J]. Journal of Liaoning University(Natural Science Edition), 2003, 30(4): 373-377. (in Chinese with English abstract)
[23]	吴文锦, 汪兰, 孙静, 等. 鸡血血红蛋白酶法提取血红素工艺优化[J]. 食品研究与开发, 2021, 42(9): 90-96.
	WU W J, WANG L, SUN J, et al. Optimization of enzymatic extraction of heme from hemoglobin in chicken blood[J]. Food Research and Development, 2021, 42(9): 90-96. (in Chinese with English abstract)
[24]	LIN B B, WANG S S, ZHOU A Q, et al. Ultrasound-assisted enzyme extraction and properties of Shatian pomelo peel polysaccharide[J]. Ultrasonics Sonochemistry, 2023, 98: 106507.
[25]	KUMCUOGLU S, YILMAZ T, TAVMAN S. Ultrasound assisted extraction of lycopene from tomato processing wastes[J]. Journal of Food Science and Technology, 2014, 51(12): 4102-4107.
[26]	HE S D, ZHANG Y, SUN H J, et al. Antioxidative peptides from proteolytic hydrolysates of false abalone (Volutharpa ampullacea perryi): characterization, identification, and molecular docking[J]. Marine Drugs, 2019, 17(2): 116.
[27]	敬思群, 张俊艳, 王德萍. 酶解-超声辅助联用提取塔尔米多糖[J]. 中国粮油学报, 2019, 34(5): 121-127.
	JING S Q, ZHANG J Y, WANG D P. Combined process of enzyme hydrolysis and ultrasound-assisted for extracting polysaccharides from Ta-er-mi[J]. Journal of the Chinese Cereals and Oils Association, 2019, 34(5): 121-127. (in Chinese with English abstract)
[28]	杨志, 李文义, 高云涛, 等. 响应面法优化针叶樱桃总黄酮的提取工艺及其抗氧化活性研究[J]. 浙江农业学报, 2020, 32(10): 1866-1872.
	YANG Z, LI W Y, GAO Y T, et al. Optimization of extraction process of total flavonoids from Acerola cherry by response surface methodology and their antioxidant activities[J]. Acta Agriculturae Zhejiangensis, 2020, 32(10): 1866-1872. (in Chinese with English abstract)
[29]	JIN Y, WANG C Y, HU W H, et al. An optimization of ultra-sonication-assisted extraction from flowers of Apocynum venetum in targeting to amount of free amino acids determined by UPLC-MS/MS[J]. Food Quality and Safety, 2019, 3(1): 52-60.
[30]	马艳弘, 孟勇, 崔晋, 等. 牛蒡多酚超声辅助酶法提取工艺及抗氧化活性[J]. 食品与生物技术学报, 2020, 39(1): 38-45.
	MA Y H, MENG Y, CUI J, et al. Optimization of ultrasonic-assisted enzyme extraction of polyphenols from burdock and its antioxidant activity evaluation[J]. Journal of Food Science and Biotechnology, 2020, 39(1): 38-45. (in Chinese with English abstract)
[31]	GUO X G, JIANG X L, ZHU Y M, et al. Unified description on principles of Fourier transform infrared spectroscopy and terahertz time-domain spectroscopy[J]. Infrared Physics & Technology, 2019, 101: 105-109.

自变量 Independent variables	不同水平(编码)的设定值Setting values for different levels (codes)
自变量 Independent variables	低 Low(-1)	中 Medium(0)	高 High(1)
(A)超声波辅助酶解时间Ultrasound-assisted enzymolytic time/h	3.5	4	4.5
(B)酶/底物浓度Enzyme/substrate concentration/(U·mg^-1)	11.25	15	18.75
(C)超声比功率Ultrasound specific power/(W·L^-1)	800	1 000	1 200

自变量 Independent variables	不同水平(编码)的设定值Setting values for different levels (codes)
自变量 Independent variables	低 Low(-1)	中 Medium(0)	高 High(1)
(A)超声波辅助酶解时间Ultrasound-assisted enzymolytic time/h	3.5	4	4.5
(B)酶/底物浓度Enzyme/substrate concentration/(U·mg^-1)	11.25	15	18.75
(C)超声比功率Ultrasound specific power/(W·L^-1)	800	1 000	1 200

序号 Number	自变量组合 Combination of independent variables	纯度Purity/%
序号 Number	自变量组合 Combination of independent variables	响应值 Response value	预测值 Predicted value	相对偏差 Relative deviation
1	A₀B₁C_-1	15.91±0.31 ab	15.92	-0.02
2	A₀B₀C₀	14.78±1.37 abc	14.13	0.65
3	A₁B_-1C₀	8.22±0.11 ghi	6.89	1.33
4	A_-1B_-1C₀	6.24±0.08 i	5.78	0.46
5	A₁B₀C₁	8.79±1.22 fgh	8.87	-0.08
6	A_-1B₁C₀	9.82±0.72 efgh	10.78	-0.96
7	A₀B_-1C₁	10.69±0.54 def	11.04	-0.36
8	A₁B₀C₁	10.52±0.22 efg	11.67	-1.16
9	A₀B_-1C_-1	8.06±1.66 hi	9.50	-1.44
10	A₀B₀C₀	16.15±0.54 a	14.13	2.02
11	A₁B₁C₀	9.91±0.86 efgh	10.00	-0.09
12	A_-1B₀C_-1	13.12±0.91 bcd	12.33	0.78
13	A₀B₁C₁	13.79±2.35 abc	12.73	1.06
14	A_-1B₀C₁	7.58±0.50 hi	7.88	-0.29
15	A₀B₀C₀	12.02±1.50 cde	14.13	-2.11
16	A₀B₀C₀	14.06±1.37 abc	14.13	-0.06
17	A₀B₀C₀	14.36±0.19 abc	14.13	0.24

序号 Number	自变量组合 Combination of independent variables	纯度Purity/%
序号 Number	自变量组合 Combination of independent variables	响应值 Response value	预测值 Predicted value	相对偏差 Relative deviation
1	A₀B₁C_-1	15.91±0.31 ab	15.92	-0.02
2	A₀B₀C₀	14.78±1.37 abc	14.13	0.65
3	A₁B_-1C₀	8.22±0.11 ghi	6.89	1.33
4	A_-1B_-1C₀	6.24±0.08 i	5.78	0.46
5	A₁B₀C₁	8.79±1.22 fgh	8.87	-0.08
6	A_-1B₁C₀	9.82±0.72 efgh	10.78	-0.96
7	A₀B_-1C₁	10.69±0.54 def	11.04	-0.36
8	A₁B₀C₁	10.52±0.22 efg	11.67	-1.16
9	A₀B_-1C_-1	8.06±1.66 hi	9.50	-1.44
10	A₀B₀C₀	16.15±0.54 a	14.13	2.02
11	A₁B₁C₀	9.91±0.86 efgh	10.00	-0.09
12	A_-1B₀C_-1	13.12±0.91 bcd	12.33	0.78
13	A₀B₁C₁	13.79±2.35 abc	12.73	1.06
14	A_-1B₀C₁	7.58±0.50 hi	7.88	-0.29
15	A₀B₀C₀	12.02±1.50 cde	14.13	-2.11
16	A₀B₀C₀	14.06±1.37 abc	14.13	-0.06
17	A₀B₀C₀	14.36±0.19 abc	14.13	0.24

项目 Item	平方和 Sum of squares	自由度 Freedom	均方值 Mean square value	F值 F value	P值 P value
模型Model	129.20	4	32.30	15.35	<0.000 1
残差Residual	25.26	12	2.10	—	—
失拟项Lack of fit	16.34	8	2.04	0.916 1	0.577 5
误差Pure error	8.92	4	2.23	—	—
总和Cor total	154.45	16	—	—	—
	R²=0.836 5	修正Corrective R²=0.782 0	—	—	—

Ultrasound-assisted enzymolytic extraction of chlorohemin from yak blood powder: response surface optimization and quality characterization

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项目 Item	系统系数 System coefficient	平方和 Sum of squares	自由度 Freedom	均方值 Mean square value	F值 F value	P值 P value
B	2.03	32.87	1	32.87	15.62	0.001 9
AC	1.82	13.18	1	13.18	6.62	0.027 8
A²	-3.94	65.44	1	65.44	31.09	<0.000 1
B²	-1.83	14.08	1	14.08	6.69	0.023 8

条件 Conditions	超声辅助酶解时间 Ultrasound-assisted enzymolytic time/h	酶/底物浓度 Enzyme/substrate concentration/(U·mg^-1)	超声比功率 Ultrasound specific power/(W·L^-1)	预测纯度 Predicted purity/%
最佳条件Optimized conditions	3.88	17.08	800.00	12.43
修正条件Modified conditions	4.00	17.00	800.00	12.38

理化指标 Physicochemical indexes		牦牛血粉 Yak blood powder	普通酶解 Enzymolysis	超声波辅助酶解 Ultrasound-assisted enzymolysis	氯化血红素标准品 Standard substance of chlorohemin
纯度Purity/%		3.54±0.03 d	7.21±0.02 c	14.89±0.13 b	95.50±0.80 a
得率Yield/%		—	27.61±0.11 b	63.30±1.32 a	—
粉体色度Powder chromaticity	L^*	43.29±1.46 a	42.72±0.93 a	34.96±0.23 c	39.74±0.51 b
	a^*	5.79±0.79 a	3.51±0.17 b	-0.54±0.13 c	0.04±0.03 c
	b^*	4.38±1.09 a	3.32±0.52 a	-0.72±0.13 b	-2.40±0.04 c
溶液色度Solution chromaticity	L^*	43.48±1.99 a	34.03±0.44 b	33.10±0.40 b	30.52±0.06 c
	a^*	17.17±0.30 a	11.12±1.31 b	7.55±0.98 c	0.15±0.03 d
	b^*	14.84±0.48 a	2.20±0.63 b	0.93±0.68 c	-1.08±0.05 d
红外光谱特征性官能团		—	700~720 cm^-1: Cl^-弯曲振动bending vibration
Characteristic functional groups		1 500~-1 700 cm^-1:—C=O伸缩振动stretching vibration
in FT-IR spectrum		2 360 & 2 900 cm^-1:—C—H弯曲振动bending vibration
		3 200~3500 cm^-1:—O—H伸缩振动stretching vibration