{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T18:35:17Z","timestamp":1775759717712,"version":"3.50.1"},"reference-count":63,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2023,8,29]],"date-time":"2023-08-29T00:00:00Z","timestamp":1693267200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"national funds","award":["SFRH\/BD\/105304\/2014"],"award-info":[{"award-number":["SFRH\/BD\/105304\/2014"]}]},{"name":"national funds","award":["UIDB\/50016\/2020"],"award-info":[{"award-number":["UIDB\/50016\/2020"]}]},{"name":"scientific collaboration of CBQF","award":["SFRH\/BD\/105304\/2014"],"award-info":[{"award-number":["SFRH\/BD\/105304\/2014"]}]},{"name":"scientific collaboration of CBQF","award":["UIDB\/50016\/2020"],"award-info":[{"award-number":["UIDB\/50016\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Hippophae rhamnoides L. leaves possess a remarkable amount of polyphenols that could serve as a natural remedy in various applications. In comparison, numerous techniques, such as conventional and high-pressure techniques, are available for extracting the bioactive fractions from sea buckthorn leaves (SBL). However, enzyme-assisted extraction (EAE) of SBL has not been comprehensively studied. The aim of this study was to optimize critical EAE parameters of SBL using the cellulolytic enzyme complex, Viscozyme L, to obtain a high-yield extract with a high concentration of bioactive compounds. In order to determine the optimal conditions for EAE, the study employed a central composite design and response surface methodology to analyze the effects of four independent factors (pH, temperature, extraction time, and enzyme concentration) on two different responses. Our findings indicated that under optimal conditions (3:15 h extraction, temperature 45 \u00b0C, pH 4.9, and 1% Viscozyme L v\/w of leaves DW), EAE yielded 28.90 g\/100 g DW of the water-soluble fraction. Furthermore, the EAE-optimized liquid extract was continuously fermented using an ancient fermentation starter, Tibetan kefir grains, which possess lactic acid bacteria (LAB) and have significant potential for use in biopreservation. Interestingly, the results indicated various potential prebiotic characteristics of LAB. Additionally, alterations in the cell wall morphology of the SBL residue after EAE were examined using scanning electron microscopy (SEM). This study significantly optimized EAE parameters for sea buckthorn leaves, providing a promising natural source of bioactive compounds for various applications, such as nutraceuticals, functional foods, and high-value products.<\/jats:p>","DOI":"10.3390\/microorganisms11092180","type":"journal-article","created":{"date-parts":[[2023,8,29]],"date-time":"2023-08-29T08:35:05Z","timestamp":1693298105000},"page":"2180","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Optimization of Enzyme-Assisted Extraction of Bioactive Compounds from Sea Buckthorn (Hippophae rhamnoides L.) Leaves: Evaluation of Mixed-Culture Fermentation"],"prefix":"10.3390","volume":"11","author":[{"given":"Viktorija","family":"Puzeryte","sequence":"first","affiliation":[{"name":"Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Kaunas, Lithuania"},{"name":"Botanical Garden, Vytautas Magnus University, Z.E. Zilibero 6, 46324 Kaunas, Lithuania"}]},{"given":"Paulina","family":"Martusevice","sequence":"additional","affiliation":[{"name":"Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Kaunas, Lithuania"},{"name":"Botanical Garden, Vytautas Magnus University, Z.E. Zilibero 6, 46324 Kaunas, Lithuania"}]},{"given":"S\u00e9rgio","family":"Sousa","sequence":"additional","affiliation":[{"name":"Universidade Cat\u00f3lica Portuguesa, CBQF-Centro de Biotecnologia e Qu\u00edmica Fina-Laborat\u00f3rio Associado, Escola Superior de Biotecnologia, Rua Diogo Botelho 1327, 4169-005 Porto, Portugal"}]},{"given":"Aiste","family":"Balciunaitiene","sequence":"additional","affiliation":[{"name":"Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Kaunas, Lithuania"},{"name":"Research Institute of Natural and Technological Sciences, Vytautas Magnus University, 40444 Kaunas, Lithuania"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1074-8384","authenticated-orcid":false,"given":"Jonas","family":"Viskelis","sequence":"additional","affiliation":[{"name":"Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Kaunas, Lithuania"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7883-2446","authenticated-orcid":false,"given":"Ana Maria","family":"Gomes","sequence":"additional","affiliation":[{"name":"Universidade Cat\u00f3lica Portuguesa, CBQF-Centro de Biotecnologia e Qu\u00edmica Fina-Laborat\u00f3rio Associado, Escola Superior de Biotecnologia, Rua Diogo Botelho 1327, 4169-005 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5546-0168","authenticated-orcid":false,"given":"Pranas","family":"Viskelis","sequence":"additional","affiliation":[{"name":"Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Kaunas, Lithuania"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5196-3948","authenticated-orcid":false,"given":"Laima","family":"Cesoniene","sequence":"additional","affiliation":[{"name":"Botanical Garden, Vytautas Magnus University, Z.E. Zilibero 6, 46324 Kaunas, Lithuania"},{"name":"Research Institute of Natural and Technological Sciences, Vytautas Magnus University, 40444 Kaunas, Lithuania"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2913-2739","authenticated-orcid":false,"given":"Dalia","family":"Urbonaviciene","sequence":"additional","affiliation":[{"name":"Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Kaunas, Lithuania"}]}],"member":"1968","published-online":{"date-parts":[[2023,8,29]]},"reference":[{"key":"ref_1","unstructured":"Li, T.S. (2004). Production et Utilisation de l\u2019Argousier (Hippophae rhamnoides L.), Presses Scientifiques du CNRC."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Letchamo, W., Ozturk, M., Altay, V., Musayev, M., Mamedov, N.A., and Hakeem, K.R. (2018). 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