Structural Study of L-Arabinose Isomerase from Latilactobacillus sakei

D-Tagatose is a rare sugar of interest as a low-calorie sweetener, and enzymatic isomerization of D-galactose is a practical production route. L-arabinose isomerase (L-AI; EC 5.3.1.4) is a promising catalyst for the above process, but many characterized L-AIs perform best at alkaline pH and high temperature and often require substantial divalent metal supplementation (e.g., Mn2+/Co2+), which complicates food-grade processing. Lactic acid bacteria (LAB) are attractive sources of food-compatible enzymes, yet structural information for LAB-derived L-AIs has been limited. Here, we report the 2.6 Å X-ray crystal structure of L-AI from Latilactobacillus sakei 23K (LsAI) and define its oligomeric assembly. Although the asymmetric unit contains a single monomer, crystallographic symmetry reconstructs a D3-symmetric homohexamer composed of two face-to-face trimers, consistent with a higher-order assembly in solution. Interface analysis shows predominantly polar interaction networks, and normalized B-factor mapping reveals localized flexibility near active-site-proximal regions. These findings provide a structural basis for understanding LAB-derived L-AIs and support structure-guided engineering toward food-grade, low-metal biocatalysts for rare-sugar production.