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In this study, controlled fermentation experiments were performed using PTMB-VIN and PTMB-GLY mixtures with varying substrate concentrations and retention times to optimize the VFA yield and production efficiency. Acid fermentation was optimized at a 3-day hydraulic retention time, and methanogenic reactions were enhanced at longer hydraulic retention times. Increasing the organic loading rate resulted in a high VFA yield (13.10\u00a0g\n                    <jats:sub>VFA-COD<\/jats:sub>\n                    . L\n                    <jats:sup>\u22121<\/jats:sup>\n                    ) and a conversion rate of approximately 40%. GLY fermentation followed the oxidative and reductive pathways at a balanced redox potential. PTMB is linked to the oxidative pathway, and GLY is involved in the reductive pathway. Tests with PTMB-GLY resulted in limiting conditions, but significant observations regarding the metabolic pathway and the effect of PTMB were made.\n                  <\/jats:p>","DOI":"10.1007\/s11356-026-37738-4","type":"journal-article","created":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T03:10:09Z","timestamp":1775877009000},"page":"6699-6711","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Insights into volatile fatty acid production and fermentation pathways of microalgae biomass combined with vinasse and glycerol"],"prefix":"10.1007","volume":"33","author":[{"given":"Dicla Ces\u00e1rio Pereira","family":"de Oliveira","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francyelle Karolynne Vieira","family":"Machado","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Agnes Adam Duarte","family":"Pinheiro","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Federico","family":"Battista","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mario Takayuki","family":"Kato","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lourdinha","family":"Florencio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5455-738X","authenticated-orcid":false,"given":"Wanderli Rog\u00e9rio Moreira","family":"Leite","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,4,11]]},"reference":[{"key":"37738_CR1","unstructured":"ANP (2023) Anu\u00e1rio Estat\u00edstico Brasileiro do Petr\u00f3leo, G\u00e1s Natural e Biocombust\u00edveis: 2023"},{"key":"37738_CR2","doi-asserted-by":"publisher","DOI":"10.1016\/J.BITEB.2022.101290","volume":"20","author":"RP Asopa","year":"2022","unstructured":"Asopa RP, Bhoi R, Saharan VK (2022) Valorization of glycerol into value-added products: a comprehensive review on biochemical route. 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