菌酶联用处理对刺参体壁降解效果及风味特性的影响

邹媛婷; 张健; 刘芳; 井月欣; 王共明; 矫春娜; 赵云苹

doi:10.13386/j.issn1002-0306.2022080279

菌酶联用处理对刺参体壁降解效果及风味特性的影响

doi: 10.13386/j.issn1002-0306.2022080279

邹媛婷^{1, 2,},
张健^2, ,,
刘芳^2, ,,
井月欣²,
王共明²,
矫春娜²,
赵云苹²

1.
上海海洋大学食品学院，上海 201306
2.
山东省海洋资源与环境研究院，山东烟台 264006

基金项目: 山东省自然科学基金重点项目——基于高品质海参肽的特殊医学用途食品等功能制品研发及产业化示范（ZR2020KC034）；山东省现代农业产业技术体系加工与质量控制岗位（SDAIT-22-07）。

详细信息

作者简介:
邹媛婷（1998−），女，硕士研究生，研究方向：海洋活性物质及功能产品，E-mail：yt3426987@163.com

通讯作者:
张健（1980−），男，博士，副研究员，研究方向：海洋活性物质及功能产品，E-mail：zjsd408@163.com

刘芳（1988−），女，硕士，工程师，研究方向：海洋活性物质及功能产品，E-mail：45637059@qq.com

中图分类号: TS254.4
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- 被引次数: 0
出版历程
- 收稿日期: 2022-08-30
- 网络出版日期: 2023-05-06
- 刊出日期: 2023-06-01

Effects of Combined Treatments of Bacteria and Enzyme on the Degradation and Flavor Components of Body Wall of Sea Cucumber

1.
College of Food Sciences & Technology, Shanghai Ocean University, Shanghai 201306, China
2.
Shandong Marine Resources and Environment Research Institute, Yantai 264006, China

摘要

摘要: 目的：为探究不同菌酶联用处理方式对刺参体壁降解效果及产物风味特性的影响。方法：采用不同处理方式：单菌（J）、单酶（M）、先菌后酶（JM）、先酶后菌（MJ）、菌酶同用（T）（菌为解淀粉芽孢杆菌，酶为复合蛋白酶）对刺参体壁进行降解，测定降解液中氨基酸组成、多肽相对分子质量分布等理化指标，同时利用气相色谱-离子迁移谱（GC-IMS）和电子舌分别测定不同处理方式下刺参体壁降解产物的挥发性和非挥发性风味成分。结果：刺参体壁经菌酶联合（JM、MJ、T）处理后，与单独作用相比，相对分子质量>1000 Da的肽含量减少了60.29%，<500 Da的肽含量增加了33.36%，联合处理下游离氨基酸总量与风味核苷酸总量的平均值较菌酶单独作用时分别增加了1.3倍和4.1倍，降解液中挥发性化合物由未处理时的43种增加到联合处理后的67种。表明刺参体壁经联合处理后风味成分变得更加复杂，联合处理可以降低小分子醛、不饱和醛等腥味成分的含量、增加酯类等愉悦性成分的含量。其中，多肽含量最高的处理组是M（34.770 mg/g），其次是JM（30.733 mg/g），感官评分最高的是JM（66.67分），其次是J（63.75分）；鲜味和甜味氨基酸之和占总氨基酸的比例中J最高，JM和M次之，T最低；菌酶联合处理中，JM处理组呈酸臭味的丙酸、刺激性气味的2-甲基丙醛相对含量较低、呈酮香味的2-戊酮、芳香味的月桂烯相对含量较高。综合各指标测定结果来看，JM降解液的风味较佳。结论：菌酶联合处理（JM、MJ、T）均能去除刺参降解产物的腥味，同时提升风味，其中JM降解综合评价最高，作为一种技术对刺参体壁进行降解，可获得具有良好风味的多肽降解产物。
- 刺参 /
- 菌酶联用 /
- 风味分析 /
- 理化分析 /
- 降解
Abstract: Objective: To investigate the effects of different combined treatments of bacteria and enzyme on the degradation and flavor components of body wall of sea cucumber. Methods: Different treatment methods: Single bacteria (J), single enzyme (M), bacteria before enzyme (JM), bacteria after enzyme (MJ), and bacteria with enzyme (T) (The bacterium was Bacillus amyloliticus and the enzyme was compound protease) were used to degrade the body wall of sea cucumber. Physical and chemical indexes such as amino acid composition and relative molecular mass distribution of polypeptides in the degradation solution were determined. Gas chromatography-ion mobility spectrometry (GC-IMS) and electronic tongue were used to determine the volatile and non-volatile flavor components of the degradation products of body wall of sea cucumber under different treatments. Results: After combined treatments of bacteria and enzyme (JM, MJ, T), compared with single action, the content of relative molecular mass of peptides larger than 1000 Da was decreased by 60.29%, while those smaller than 500 Da was increased by 33.36%. The content of total free amino acid and nucleotide of combined treatments were increased by 1.3 and 4.1 times, respectively, compared with that of bacterial enzyme alone. The volatile compounds in the degradation solution increased from 43 without treatment to 67 after combined treatments, indicating that the flavor components of body wall of sea cucumber became more complex after combined treatment. Combined treatment could reduce the content of small molecular aldehydes, unsaturated aldehydes and other fishy ingredients, increase the content of esters and other pleasant ingredients. Among them, the treatment group with the highest polypeptide content was M (34.770 mg/g), followed by JM (30.733 mg/g), and the sensory score was highest in JM (66.67), followed by J (63.75). In the proportion of the sum of umami and sweet amino acids to the total amino acids, J was the highest, followed by JM and M, and T was the lowest. In the combined treatments of bacterial and enzyme, the relative contents of propionic acid with sour odor and 2-methylpropionic aldehyde with pungent odor were lower, and the relative contents of 2-pentanone with ketone flavor and laurene with aromatic flavor were higher in the JM treatment group. According to the results of each index, the flavor of JM degradation solution was better. Conclusion: The combined treatments of bacteria and enzyme (JM, MJ, T) can remove the fishy taste of the degradation products of sea cucumber, and improve the aromatic flavor at the same time. JM has the highest evaluation of degradation. As a technology to degrade the body wall of sea cucumber, polypeptide degradation products with good flavor can be obtained.
- Stichopus japonicus /
- bacterial enzyme combination /
- flavor analysis /
- physicochemical analysis /
- degradation

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图 1 不同菌对发酵液多肽含量和感官评分的影响

Figure 1. Effects of different bacteria on polypeptide content and sensory score of fermentation broth

注：同一指标不同字母表示差异显著（P<0.05），图2、图3同。

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图 2 不同蛋白酶对酶解液多肽含量和感官评分的影响

Figure 2. Effects of different proteases on polypeptide content and sensory score of enzymatic hydrolysate

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图 3 不同处理方式对降解液多肽含量、感官评分和游离氨基酸总量的影响

Figure 3. Effects of different treatment methods on polypeptide content, sensory score and total free amino acid in degradation solution

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图 4 不同处理方式降解液挥发性化合物的Gallery Plot图

Figure 4. Gallery Plot of volatile compounds in different treatment degradation solutions

注：从上往下依次为Y、JM、T、MJ、M、J样品；图中每个亮点代表一种挥发性化合物，每一行代表一个样品中选取的全部信号峰，数字编号的是未鉴定出的物质，其中：A为Y的挥发性成分；B为M的挥发性成分；C为菌酶共同处理的挥发性成分；D为J的挥发性成分。

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图 5 不同处理方式降解液的PCA图

Figure 5. PCA diagram of degradation solution with different treatment methods

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图 6 不同处理方式降解液的聚类分析图

Figure 6. Cluster analysis diagram of degradation solution with different treatment methods

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表 1 不同菌种的发酵条件

Table 1. Fermentation conditions of different strains

菌种种类	pH	温度（℃）	时间（h）
枯草芽孢杆菌	7.0	37	48
植物乳杆菌	6.8	37	48
肠膜明串珠菌	7.0	37	48
解淀粉芽孢杆菌	7.2	30	48
鲁氏酵母	7.0	28	48

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表 2 不同酶制剂的酶解条件

Table 2. Enzymatic hydrolysis conditions of different enzyme preparations

蛋白酶种类	pH	温度（℃）	时间（h）
风味蛋白酶	7.5	50	4
复合蛋白酶	6.0	50	4
木瓜蛋白酶	6.5	55	4
碱性蛋白酶	8.0	50	4

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表 3 感官评分标准

Table 3. Sensory rating criteria

项目	评分标准
项目	1~2分	3~5分	6~8分	9~10分
腥味（30分）	腥味较重	腥味中等	腥味较淡	基本无腥味
气味（30分）	发酵味、酸味过重并伴随一定的异味	具有刺参固有的气味，无不良气味	具有刺参固有的气味，发酵产生的鲜香味	具有刺参固有的气味，令人愉悦的果香味和清爽的酸味
滋味（40分）	口感较差，过苦、过咸、有异味	口感一般，偏苦、偏咸	口感适宜，微甜、微咸	口感柔和，酸甜适宜、鲜香可口

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表 4 不同处理方式降解液的多肽相对分子质量分布

Table 4. Relative molecular mass distribution of peptides in degradation solution with different treatment methods

相对分子质量（Da）	J（%）	M（%）	JM（%）	MJ（%）	T（%）
>3000	5.921	0.051	0.109	0.091	0
2000~3000	8.885	0.813	1.175	1.297	0.11
1000~2000	19.445	9.575	10.324	9.919	3.592
500~1000	21.956	28.908	25.555	22.013	16.886
<500	43.794	60.653	62.837	66.680	79.412

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表 5 不同处理方式降解液的氨基酸组成

Table 5. Amino acid composition of degradation solution by different treatment methods

序号	氨基酸	含量（g/100 g干重）
序号	氨基酸	Y	J	M	JM	MJ	T
1	天门冬氨酸Asp	6.04±0.04^ab	4.99±0.11^d	6.37±0.11^a	5.78±0.25^bc	5.67±0.14^c	5.10±0.11^d
2	苏氨酸Thr	3.13±0.02^b	2.23±0.08^d	3.35±0.04^a	2.56±0.11^c	1.98±0.05^e	1.59±0.03^f
3	丝氨酸Ser	3.08±0.03^a	2.19±0.04^c	3.18±0.04^a	2.55±0.1^b	2.20±0.05^c	1.81±0.03^d
4	谷氨酸Glu	9.82±0.11^a	7.34±0.15^c	10.01±0.15^a	8.49±0.36^b	7.35±0.17^c	6.2±0.13^d
5	甘氨酸Gly	10.32±0.14^a	8.15±0.23^d	9.65±0.18^b	8.98±0.36^c	8.6±0.17^cd	7.43±0.16^e
6	丙氨酸Ala	4.39±0.04^a	2.98±0.03^c	4.29±0.09^a	3.48±0.14^b	2.77±0.07^d	2.22±0.05^e
7	半胱氨酸Cys	1.39±0.09^ab	1.50±0.18^a	1.49±0.04^ab	1.42±0.02^ab	1.40±0.03^ab	1.26±0.03^b
8	缬氨酸Val	2.28±0.02^b	1.84±0.03^e	2.52±0.01^a	2.14±0.06^c	2.11±0.04^c	1.97±0.05^d
9	蛋氨酸Met	0.86±0.01^bc	0.56±0.01^cd	0.46±0.02^d	0.98±0.28^b	1.37±0.10^a	1.11±0.14^ab
10	异亮氨酸Ile	1.74±0.01^b	1.23±0.04^d	1.85±0.06^a	1.45±0.05^c	1.23±0.02^d	1.19±0.03^d
11	亮氨酸Leu	2.63±0.01^b	2.04±0.01^d	3.05±0.03^a	2.45±0.10^c	2.68±0.06^b	2.57±0.05^bc
12	酪氨酸Tyr	2.01±0.02^bc	1.91±0.07^c	2.35±0.02^a	2.27±0.04^ab	2.12±0.16^abc	2.01±0.23^bc
13	苯丙氨酸Phe	1.45±0.01^c	1.41±0.03^c	1.75±0.04^ab	1.72±0.07^b	1.82±0.01^a	1.73±0.04^ab
14	赖氨酸Lys	1.88±0.01^c	1.80±0^c	2.25±0.04^a	2.16±0.08^ab	2.19±0.03^ab	2.12±0.05^b
15	脯氨酸Pro	4.87±0.05^a	3.77±0.05^c	4.74±0.05^a	4.27±0.13^b	3.78±0.08^c	3.29±0.08^d
16	组氨酸His	0.60±0^b	0.46±0.01^c	0.74±0.01^a	0.55±0.04^b	0.41±0.04^c	0.44±0.02^c
17	精氨酸Arg	4.74±0.08^a	3.07±0.07^c	4.58±0.08^a	3.54±0.18^b	2.74±0.04^d	2.12±0.04^e
18	TAA	61.22±0.59^a	47.48±0.92^d	62.61±0.8^a	54.78±1.77^b	50.43±0.83^c	44.19±0.93^e
19	DAA	15.86±0.18^a	12.33±0.32^c	16.38±0.32^a	14.27±0.76^b	13.02±0.37^c	11.30±0.29^d
20	SAA	25.79±0.35^a	19.33±0.55^c	25.2±0.49^a	21.84±1.03^b	19.33±0.51^c	16.35±0.43^d
21	BAA	18.18±0.07^b	14.33±0.03^f	19.55±0.18^a	17.25±0.34^c	16.68±0.08^d	15.28±0.38^e
22	DAA+SAA	41.65±0.53^a	31.65±0.87^c	41.58±0.81^a	36.11±1.78^b	32.35±0.89^c	27.65±0.72^d
23	（DAA+SAA）/TAA	0.68±0^a	0.67±0^b	0.66±0^b	0.66±0.01^b	0.64±0^c	0.63±0^d
24	BAA/TAA	0.30±0^d	0.30±0.01^cd	0.31±0^c	0.32±0^c	0.33±0^b	0.35±0^a
注：TAA为总氨基酸；DAA为鲜味氨基酸（谷氨酸、天冬氨酸）；SAA为甜味氨基酸（甘氨酸、丙氨酸、丝氨酸、脯氨酸、苏氨酸）；BAA为苦味氨基酸（酪氨酸、苯丙氨酸、缬氨酸、组氨酸、异亮氨酸、亮氨酸、赖氨酸、精氨酸、甲硫氨酸）；同行不同小写字母表示不同处理组差异显著（P<0.05）。

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表 6 不同处理方式降解液的核苷酸含量

Table 6. Nucleotide content of degradation solution by different treatment methods

不同处理方式	鲜味物质含量（mg/100 g）
不同处理方式	5’-AMP （腺苷酸）	5’-GMP （鸟苷酸）	5’-IMP （肌苷酸）	风味核苷酸
Y	0.43±0.00^e	−	−	0.43±0.00^e
J	2.17±0.06^c	11.33±0.78^b	−	13.50±0.84^c
M	0.76±0.00^d	−	−	0.76±0.00^d
JM	2.45±0.07^a	11.31±0.78^b	−	13.76±0.85^c
MJ	2.29±0.04^b	32.90±1.36^a	−	35.19±1.4^b
T	2.20±0.02^bc	36.39±2.31^a	−	38.59±2.33^a
注：−表示未检出；风味核苷酸为5’-GMP、5’-IMP和5’-AMP之和；同列小写字母不同表示不同处理组差异显著（P<0.05）。

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