Authentication of Suzhou Yangcheng Lake Hairy Crab Based on Protein Fingerprint

doi:10.13304/j.nykjdb.2023.0115

Abstract

Abstract:

Crab chelate muscle tissue protein is rich in species and appears in the mass spectrum with different intensity peaks. Crab chelate muscle tissue protein is extracted， and the characteristic index of crab chelate muscle tissue protein spectrum was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS） technology， and the characteristic index is found to achieve the qualitative and quantitative detection of crab chelate muscle tissue protein. In this study， protein molecular fingerprint databases of different categories of crabs were established by analyzing the histoprotein MALDI-TOF MS technology， and the database was deeply analyzed with machine deep learning algorithm to establish the homology analysis method of Yangcheng Lake hairy crabs. The results showed that the protein profiles of hairy crabs in different growing environments showed different expressions.There was an obvious difference when m/z（mass-charge ratio，m/z） was less than 10 000，the scanning results showed that the low m/z region （<10 000） had higher peak intensity and higher sub-marking rate，this region could be used as the mass spectrum resolution region to distinguish Yangcheng Lake hairy crabs from other lake crabs. Above results showed MALDI-TOF MS technique could be used to analyze the different protein expressions of hairy crabs from different places， which could play a role in identifying hairy crab from different places.

Key words: MALDI-TOF mass spectrometry, Yangcheng Lake hairy crab, homology analysis

摘要：

蟹螯肌肉组织蛋白质种类丰富，以不同强度的质谱峰出现在质谱图中，提取蟹鳌肌肉组织蛋白，利用基质辅助激光解析电离飞行时间质谱（matrix-assisted laser desorption/ionization time-of-flight mass spectrometry，MALDI-TOF MS）技术，对蟹鳌肌肉组织蛋白质谱进行特征指标分析，找到特征指标，可实现蟹鳌肌肉组织蛋白定性定量检测。通过分析蟹鳌组织蛋白的MALDI-TOF MS图谱规律，解析不同品类螃蟹蛋白质分子指纹图谱，并配合机器深度学习算法对数据库进行深度分析，建立阳澄湖大闸蟹的同源性分析方法。结果表明，不同生长环境的大闸蟹蟹鳌组织蛋白质谱图有不同的表达。m/z（质荷比，mass-to-charge ratio）范围在10 000内均有明显差异，扫描结果显示，低质量区（m/z<10 000）出峰强度较高且分辨率较高，该区域可以作为区分阳澄湖大闸蟹与其他类别湖蟹的质谱分辨区。以上结果表明，可以通过MALDI-TOF MS技术对不同产地大闸蟹的差异性蛋白质的表达进行分析，从而鉴别大闸蟹的产地。

关键词: MALDI-TOF质谱定量技术, 阳澄湖大闸蟹, 同源性分析

CLC Number:

S963

Yahan ZHANG, Youyong ZHANG, Dan YUAN, Min XU, Yezhong CAO. Authentication of Suzhou Yangcheng Lake Hairy Crab Based on Protein Fingerprint[J]. Journal of Agricultural Science and Technology, 2023, 25(11): 80-89.

张雅涵, 张有勇, 袁丹, 徐敏, 曹叶中. 基于蛋白指纹图谱的阳澄湖大闸蟹真伪鉴别研究[J]. 中国农业科技导报, 2023, 25(11): 80-89.

Figures/Tables 10

Fig. 1 Tissue of the claw muscleNote： A—The first crab foot；B—The second crab foot；C—The third crab foot；D—The fourth crab foot.

Fig. 2 Appearance of different hairy crabsA：Non-Yangcheng Lake hairy crab abdomen；B：Yangcheng Lake hairy crab abdomen；C：Non-Yangcheng Lake hairy crab back；D：Yangcheng Lake hairy crab back

Fig. 3 3D anatomy and coronal image of different hairy crabsA：3D anatomy of Yangcheng Lake hairy crab（female crab）；B：3D anatomy of Yangcheng Lake hairy crab（male crab）；C：Coronal image of Non-Yangcheng Lake hairy crab（female crab）；D：Coronal image of Non-Yangcheng Lake hairy crab（male crab）

Fig. 4 Mass spectra of different hairy crabs collected by MALDI-TOF MSA：Yangcheng Lake hairy crab；B：Non-Yangcheng Lake hairy crab

Fig. 5 Comparison of mass spectra differences of different hairy crabs collected by MALDI-TOF MSA：Yangcheng Lake hairy crab；B：Non-Yangcheng Lake hairy crab

Fig. 6 Comparison of mass spectra of hairy crabs under different m/zA： Yangcheng Lake hairy crab， m/z<10 000； B： Yangcheng Lake hairy crab， m/z>10 000； C： Non-Yangcheng Lake hairy crab， m/z<10 000； D： Non-Yangcheng Lake hairy crab， m/z>10 000

Fig. 7 Comparison of mass spectra of hairy crabs under laser energy 9 μJA：Yangcheng Lake hairy crab；B：Non-Yangcheng Lake hairy crab；R is resolution

Fig. 8 Result of SVMNote： A—Distribution of of Yangcheng Lake hairy crab；B—Distribution of of Non-Yangcheng Lake hairy crab。

Table 1 Sample statistics

组别

Group

平均值

Average value

数量

Number

标准偏差 Standard deviation

标准误差平均值

Mean standard error

阳澄湖大闸蟹

Yangcheng Lake hairy crab

10 621.27

2 551.009

444.074

非阳澄湖大闸蟹

Non-Yangcheng Lake hairy crab

10 381.76

2 357.335

410.359

Table 2 Test of samples

组别

Group

配对差值

Pairing difference

t值

t value

自由度 Degree of freedom

P值

P value

平均值

Average value

标准偏差Standard deviation

标准误差平均值 Mean standard

error

差值的95%置信区间

P5% confidence interval for the difference

下限

Lower limit

上限

Upper limit

阳澄湖大闸蟹 Yangcheng Lake hairy crab

非阳澄湖大闸蟹

Non-Yangcheng Lake hairy crab

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