Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted Extraction

Thai fermented soybeans (TFSs) contain phytochemicals with anti-diabetic benefits. In this study, an initial non-optimized TFS extract (TFSE) was prepared using a conventional triplicate 80% ethanol extraction method and evaluated for its biological activity. TFSE effectively reversed TNF-α-induced...

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Main Authors: Piya Temviriyanukul, Woorawee Inthachat, Ararat Jaiaree, Jirarat Karinchai, Pensiri Buacheen, Supachai Yodkeeree, Tanongsak Laowanitwattana, Teera Chewonarin, Uthaiwan Suttisansanee, Arisa Imsumran, Ariyaphong Wongnoppavich, Pornsiri Pitchakarn
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Foods
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Online Access:https://www.mdpi.com/2304-8158/14/15/2603
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author Piya Temviriyanukul
Woorawee Inthachat
Ararat Jaiaree
Jirarat Karinchai
Pensiri Buacheen
Supachai Yodkeeree
Tanongsak Laowanitwattana
Teera Chewonarin
Uthaiwan Suttisansanee
Arisa Imsumran
Ariyaphong Wongnoppavich
Pornsiri Pitchakarn
author_facet Piya Temviriyanukul
Woorawee Inthachat
Ararat Jaiaree
Jirarat Karinchai
Pensiri Buacheen
Supachai Yodkeeree
Tanongsak Laowanitwattana
Teera Chewonarin
Uthaiwan Suttisansanee
Arisa Imsumran
Ariyaphong Wongnoppavich
Pornsiri Pitchakarn
author_sort Piya Temviriyanukul
collection DOAJ
description Thai fermented soybeans (TFSs) contain phytochemicals with anti-diabetic benefits. In this study, an initial non-optimized TFS extract (TFSE) was prepared using a conventional triplicate 80% ethanol extraction method and evaluated for its biological activity. TFSE effectively reversed TNF-α-induced insulin resistance in 3T3-L1 adipocytes by enhancing insulin-stimulated glucose uptake, indicating anti-diabetic potential. TFSE also upregulated the phosphorylation of AKT (a key insulin signaling mediator) and the expression of adipogenic proteins (PPARγ, CEBPα) in TNF-α-exposed 3T3-L1, suggesting the mitigation of adipocyte dysfunction; however, the results did not reach statistical significance. The conventional extraction process was labor-intensive and time-consuming, and to enhance extraction efficiency and bioactivity, the process was subsequently optimized using environmentally friendly microwave-assisted extraction (MAE) in combination with the Box–Behnken design (BBD) and response surface methodology (RSM). The optimized extract (O-TFSE) was obtained over a significantly shorter extraction time and exhibited higher levels of total flavonoids and antioxidant activity in comparison to TFSE, while showing reduced levels of isoflavones (daidzein, genistein, and glycitein) in relation to TFSE. Interestingly, O-TFSE retained similar efficacy in reversing TNF-α-induced insulin resistance and demonstrated significantly stronger α-glucosidase and α-amylase inhibitory activities, indicating its enhanced potential for diabetes management. These results support the use of MAE as an efficient method for extracting functional compounds from TFS for functional foods targeting insulin resistance and type 2 diabetes mellitus.
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spelling doaj-art-790c7ada4054463d9e4b5c8fa02a4ad72025-08-20T03:02:55ZengMDPI AGFoods2304-81582025-07-011415260310.3390/foods14152603Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted ExtractionPiya Temviriyanukul0Woorawee Inthachat1Ararat Jaiaree2Jirarat Karinchai3Pensiri Buacheen4Supachai Yodkeeree5Tanongsak Laowanitwattana6Teera Chewonarin7Uthaiwan Suttisansanee8Arisa Imsumran9Ariyaphong Wongnoppavich10Pornsiri Pitchakarn11Institute of Nutrition, Mahidol University, Salaya, Nakhon Pathom 73170, ThailandInstitute of Nutrition, Mahidol University, Salaya, Nakhon Pathom 73170, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandInstitute of Nutrition, Mahidol University, Salaya, Nakhon Pathom 73170, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandDepartment of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, ThailandThai fermented soybeans (TFSs) contain phytochemicals with anti-diabetic benefits. In this study, an initial non-optimized TFS extract (TFSE) was prepared using a conventional triplicate 80% ethanol extraction method and evaluated for its biological activity. TFSE effectively reversed TNF-α-induced insulin resistance in 3T3-L1 adipocytes by enhancing insulin-stimulated glucose uptake, indicating anti-diabetic potential. TFSE also upregulated the phosphorylation of AKT (a key insulin signaling mediator) and the expression of adipogenic proteins (PPARγ, CEBPα) in TNF-α-exposed 3T3-L1, suggesting the mitigation of adipocyte dysfunction; however, the results did not reach statistical significance. The conventional extraction process was labor-intensive and time-consuming, and to enhance extraction efficiency and bioactivity, the process was subsequently optimized using environmentally friendly microwave-assisted extraction (MAE) in combination with the Box–Behnken design (BBD) and response surface methodology (RSM). The optimized extract (O-TFSE) was obtained over a significantly shorter extraction time and exhibited higher levels of total flavonoids and antioxidant activity in comparison to TFSE, while showing reduced levels of isoflavones (daidzein, genistein, and glycitein) in relation to TFSE. Interestingly, O-TFSE retained similar efficacy in reversing TNF-α-induced insulin resistance and demonstrated significantly stronger α-glucosidase and α-amylase inhibitory activities, indicating its enhanced potential for diabetes management. These results support the use of MAE as an efficient method for extracting functional compounds from TFS for functional foods targeting insulin resistance and type 2 diabetes mellitus.https://www.mdpi.com/2304-8158/14/15/2603Thua-Naoinsulin resistancediabetesα-glucosidaseα-amylaseisoflavones
spellingShingle Piya Temviriyanukul
Woorawee Inthachat
Ararat Jaiaree
Jirarat Karinchai
Pensiri Buacheen
Supachai Yodkeeree
Tanongsak Laowanitwattana
Teera Chewonarin
Uthaiwan Suttisansanee
Arisa Imsumran
Ariyaphong Wongnoppavich
Pornsiri Pitchakarn
Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted Extraction
Foods
Thua-Nao
insulin resistance
diabetes
α-glucosidase
α-amylase
isoflavones
title Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted Extraction
title_full Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted Extraction
title_fullStr Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted Extraction
title_full_unstemmed Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted Extraction
title_short Enhancement of Phytochemicals and Antioxidant Activity of Thai Fermented Soybean Using Box–Behnken Design Guided Microwave-Assisted Extraction
title_sort enhancement of phytochemicals and antioxidant activity of thai fermented soybean using box behnken design guided microwave assisted extraction
topic Thua-Nao
insulin resistance
diabetes
α-glucosidase
α-amylase
isoflavones
url https://www.mdpi.com/2304-8158/14/15/2603
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