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This research was designed to optimize IND production at varying growth conditions, to assess its chemical stability, and to study its possible biological activities associated with diabetes, obesity, and insulin resistance. Medium types played an important role in both BL21\u2010DE3 cell growth and IND production. The BL21\u2010DE3 cell growth was maximum in PGB medium (2.76\u2009g\/L\/24\u2009h), then in LB\u2010O medium (1.29\u2009g\/L\/24\u2009h), LB medium (1.14\u2009g\/L\/24\u2009h), and NB medium (0.94\u2009g\/L\/24\u2009h). The IND production was 75.8, 49.8, and 38.9\u2009mg\/L\/24\u2009h in LB, NB, and PGB media, respectively, while the experimental maximum IND production was achieved in LB\u2010O medium (177.4\u2009mg\/L\/24\u2009h), which closely resembled the theoretical IND production of 180\u2009mg\/L\/24\u2009h. IND exhibited high cellular absorption properties, low cytotoxicity, and dose\u2010dependent inhibition of \u03b1\u2010amylase and lipase enzymes at IC\n                    <jats:sub>50<\/jats:sub>\n                    values of 0.69 and 0.37\u2009\u00b5g\/mL, respectively. Furthermore, IND enhanced 5\u2032\u2010adenosine monophosphate\u2010activated protein kinase\u00a0(AMPK) and protein kinase B (AKT) signaling pathways at EC\n                    <jats:sub>50<\/jats:sub>\n                    values of 1.36 and 4.59\u2009\u00b5g\/mL, respectively. The binding affinity of IND was also observed towards \u03b1\u2010amylase, lipase, AMPK, and AKT enzymes via molecular docking with binding energies (BE) of \u22127.4, \u22128.1, \u22126.9, and \u22125.5\u2009kcal\/mol, respectively. All these findings were confirmed through human spermatozoa culture, where IND stimulated glucose metabolism and AMPK and AKT signaling pathways.\n                  <\/jats:p>","DOI":"10.1002\/mbo3.70429","type":"journal-article","created":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T16:18:00Z","timestamp":1791389880000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Sustainable Overproduction of the Bacterial Pigment Indigoidine as a Natural Colorant: Stability Evaluation, Inhibition of Obesity\u2010Diabetes\u2010Related Digestive Enzymes, and Activation of AMPK\u2013AKT"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-4296-4038","authenticated-orcid":false,"given":"Tahani Ali","family":"Asseri","sequence":"first","affiliation":[{"name":"Department of Biology, College of Science King Khalid University Abha Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,7]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2025.118277"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2025.118544"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fbio.2025.107122"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fbio.2025.107356"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biteb.2026.102741"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.3390\/medicina61040684"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1996.tb01045.x"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/mbo3.70282"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.4103\/aja2025108"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-19171-4"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2019.103605"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.dyepig.2026.113881"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biortech.2025.132808"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/biology12060827"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00297-12"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.2174\/1566524020666201203164910"},{"key":"e_1_2_11_18_1","first-page":"355","volume-title":"Current Developments in Biotechnology and Bioengineering","author":"Dubey K. 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