The Study of Alanine Transaminase Activity in Tissues of Silkworm (Bombyx mori) via Direct Analysis in Real-Time (DART) Mass Spectrometry

Author:

Wu Guohua12ORCID,Jiang Lei2,Guo Jianjun3,Li Wushuang1,Ma Lin1ORCID,Tang Bozhi1,Liu Charles C.4

Affiliation:

1. College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China

2. College of Environmental and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212100, China

3. College of Agriculture, Anshun University, Anshun 561000, China

4. ASPEC Technologies Limited, Beijing 100101, China

Abstract

Alanine transaminase (ALT) is an important amino acid-metabolizing enzyme in silkworm Bombyx mori L., and is mainly involved in transferring glutamate to alanine (serving as an essential precursor in silk protein synthesis) through transamination. Therefore, it is generally believed that silk protein synthesis in the silk gland and the cocoon quantity increase with the increase in ALT activity to a certain extent. Here, a novel analytical method was developed to determine the ALT activity in several key tissues of Bombyx mori L. including the posterior silk gland, midgut, fat body, middle silk gland, trachea and hemolymph, by combining the direct-analysis-in-real-time (DART) ion source with a triple-quadrupole mass spectrometer. In addition, a traditional ALT activity assay, the Reitman–Frankel method, was also used to measure ALT activity for comparison. The ALT activity results obtained via the DART-MS method are in good agreement with those obtained via the Reitman–Frankel method. However, the present DART-MS method provides a more convenient, rapid and environmentally friendly quantitative method for ALT measurement. Especially, this method can also monitor ALT activity in different tissues of Bombyx mori L. in real time.

Funder

National Project of Risk Assessment for Quality and Safety of Special Agro-products

Publisher

MDPI AG

Subject

Chemistry (miscellaneous),Analytical Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Molecular Medicine,Drug Discovery,Pharmaceutical Science

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