Belghit Ikram, Lock Erik-Jan, Fumière Olivier, Lecrenier Marie-Caroline, Renard Patricia, Dieu Marc, Berntssen Marc H G, Palmblad Magnus, Rasinger Josef D
Institute of Marine Research, P.O. Box 1870 Nordnes, 5817 Bergen, Norway.
Centre Wallon de Recherches agronomiques (CRA-W), 5030 Gembloux, Belgium.
Animals (Basel). 2019 May 7;9(5):222. doi: 10.3390/ani9050222.
Insect protein has the potential to become a sustainable feed ingredient for the rapidly growing aquaculture industry. In the European Union, insect derived protein is placed under the same legislation as processed animal proteins (PAP). It is therefore of interest to develop methods for regulatory use, which unambiguously identify the species origin of insect-based ingredients. We performed (i) total protein quantification of insect samples using the traditional nitrogen-to-protein conversion factor of 6.25 and the sum of anhydrous amino acids, (ii) quantitative amino acid profiling and (iii) high-throughput tandem mass spectrometry to describe and differentiate 18 different commercial-grade insect meal samples derived from (8), (5), (3) and (2). In addition, we investigated and compared different protein extraction and digestion protocols for proteomic analysis. We found that irrespective of sample preparation, shotgun proteomics in combination with direct spectral comparison were able to differentiate insect meal according to their taxonomic classification. The insect specific spectral libraries created in the present work can in future be used to develop more sensitive targeted methods of insect PAP identification and quantification in commercial feed mixtures.
昆虫蛋白有潜力成为快速发展的水产养殖业可持续的饲料成分。在欧盟，昆虫衍生蛋白与加工动物蛋白（PAP）受同一法规监管。因此，开发用于监管的方法很有必要，这些方法要能明确鉴定基于昆虫的成分的物种来源。我们进行了以下操作：（i）使用传统的6.25氮到蛋白转换因子和无水氨基酸总和对昆虫样本进行总蛋白定量，（ii）进行定量氨基酸分析，（iii）使用高通量串联质谱来描述和区分来自（8种）、（5种）、（3种）和（2种）的18种不同商业级昆虫粉样本。此外，我们研究并比较了蛋白质组学分析中不同的蛋白质提取和消化方案。我们发现，无论样本制备如何，鸟枪法蛋白质组学结合直接光谱比较能够根据昆虫粉的分类学分类进行区分。本研究中创建的昆虫特异性光谱库未来可用于开发更灵敏的靶向方法，以鉴定和定量商业饲料混合物中的昆虫PAP。
