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This study introduces a significant advancement in PLP biosynthesis by enhancing the stability and activity of <jats:italic>Escherichia coli<\/jats:italic>\u2013derived pyridoxal kinase (<jats:italic>Ec<\/jats:italic>PdxK) through immobilization on an innovative epoxy resin, LXTE\u2010600. Our approach involved the systematic optimization of enzyme loading, coupling duration, and temperature, which resulted in improved immobilization efficiency and a high loading capacity of 80\u00a0mg\/g. The characterization of immobilized <jats:italic>Ec<\/jats:italic>PdxK@LXTE\u2010600 was conducted using Fourier transform infrared spectroscopy (FTIR) and confocal laser scanning microscopy (CLSM), confirming successful immobilization. This process notably enhanced the enzyme's performance, increasing its tolerance to pH and temperature fluctuations, thereby improving its thermal stability. The immobilized <jats:italic>Ec<\/jats:italic>PdxK@LXTE\u2010600 retained over 80% of its initial activity after 4\u00a0weeks of storage at 4\u00b0C and could be reused up to eight cycles while maintaining more than 70% of its initial activity. These findings not only demonstrate the efficacy of the LXTE\u2010600\u2010based immobilization method but also suggest promising industrial applications for the sustainable production of PLP, potentially revolutionizing approaches in biotechnological and pharmaceutical sectors.<\/jats:p>","DOI":"10.1002\/bab.2729","type":"journal-article","created":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T01:19:56Z","timestamp":1738631996000},"page":"1208-1221","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Advancing PLP Biosynthesis: Enhanced Stability and Activity of <i>Ec<\/i>PdxK via LXTE\u2010600 Immobilization"],"prefix":"10.1002","volume":"72","author":[{"given":"Yunhui","family":"Ye","sequence":"first","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences Nanjing University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heng","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences Nanjing University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Fan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences Nanjing University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qilong","family":"Yao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences Nanjing University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenhong","family":"Lu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences Nanjing University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junzhong","family":"Liu","sequence":"additional","affiliation":[{"name":"Nanjing Institute for Comprehensive Utilization of Wild Plants CHINA CO\u2010OP  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-3354-0667","authenticated-orcid":false,"given":"Qingcai","family":"Jiao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences Nanjing University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,2,3]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253\u2010021\u201011199\u2010w"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00394\u2010022\u201003033\u20104"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.5483\/BMBRep.2022.55.9.090"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja411785r"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.embor.embor914"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1107\/S1399004713031283"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja9907616"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.6b03056"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.48.30583"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M305967200"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2011.02.003"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.2174\/1381612811319310007"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.14599"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules14010329"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2019.00032"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.3891\/acta.chem.scand.09\u20100001"},{"issue":"7","key":"e_1_2_8_18_1","first-page":"2705","article-title":"A New Synthesis of Pyridoxine (Vitamin B6)","volume":"27","author":"Harris E. 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