Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory Peptides

The study aimed to establish the effective extraction methods for Chlamydomonas reinhardtii proteins and screen peptide segments with inhibitory effects on monoamine oxidase A (MAO-A). The proteins were extracted, separated, and purified by swelling, ultrasonication, alkaline dissolution, and saltin...

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Main Authors: Meiting LIU, Ya MAO, Jintao XIE, Wenxia LIU, Lixia WU, Yishan WU, Mei WANG, Xuewu ZHANG
Format: Article
Language:zho
Published: The editorial department of Science and Technology of Food Industry 2025-06-01
Series:Shipin gongye ke-ji
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Online Access:http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024070176
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author Meiting LIU
Ya MAO
Jintao XIE
Wenxia LIU
Lixia WU
Yishan WU
Mei WANG
Xuewu ZHANG
author_facet Meiting LIU
Ya MAO
Jintao XIE
Wenxia LIU
Lixia WU
Yishan WU
Mei WANG
Xuewu ZHANG
author_sort Meiting LIU
collection DOAJ
description The study aimed to establish the effective extraction methods for Chlamydomonas reinhardtii proteins and screen peptide segments with inhibitory effects on monoamine oxidase A (MAO-A). The proteins were extracted, separated, and purified by swelling, ultrasonication, alkaline dissolution, and salting-out. Subsequently, the polypeptide content, amino acid composition, molecular weight distribution and the MAO-A inhibitory effect of proteins treated with enzymatic hydrolysis and ultrafiltration were determined. Peptide sequences with high inhibitory potential were selected by bioinformatics tools and molecular docking with MAO-A was applied to analyze the binding sites. The results indicated that a protein extraction yield of 50.98%±2.50% was achieved through swelling for 4 h, followed by four cycles of 240 W ultrasonication (10 min per cycle), alkaline dissolution at pH9.5, acid precipitation at pH3.5, and the treatment with saturated ammonium sulfate at a ratio of 1:2. Notably, the alkaline protease hydrolysate (APH) and neutral protease hydrolysate (NPH) exhibited high inhibitory effects on MAO-A (P<0.05), with IC50 values of 3.437 and 2.699 mg/mL, respectively. The fractions with molecular weights less than 3 kDa from APH and NPH (APH-Ⅲ and NPH-Ⅲ) displayed significantly higher MAO-A inhibitory effect (P<0.05), with IC50 values of 3.116 and 0.909 mg/mL, respectively. Molecular docking results showed that two peptides, ASDDAVHGWGGPGGY and AYSGETQSEGGFAPNR from NPH-Ⅲ, could effectively bind to the active sites His488, Glu286, Tyr53, Tyr268, Asp343 and Glu485 of MAO-A.
format Article
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issn 1002-0306
language zho
publishDate 2025-06-01
publisher The editorial department of Science and Technology of Food Industry
record_format Article
series Shipin gongye ke-ji
spelling doaj-art-af1cf223ef104956bd14e28acd3b09b12025-08-20T02:08:38ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062025-06-01461222623610.13386/j.issn1002-0306.20240701762024070176-12Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory PeptidesMeiting LIU0Ya MAO1Jintao XIE2Wenxia LIU3Lixia WU4Yishan WU5Mei WANG6Xuewu ZHANG7College of Food Science and Engineering, South China University of Technology, Guangzhou 510641, ChinaZhanjiang Institute of Supervision and Test on Quality & Measurement, Zhanjiang 524094, ChinaZhanjiang Institute of Supervision and Test on Quality & Measurement, Zhanjiang 524094, ChinaZhanjiang Institute of Supervision and Test on Quality & Measurement, Zhanjiang 524094, ChinaZhanjiang Institute of Supervision and Test on Quality & Measurement, Zhanjiang 524094, ChinaZhanjiang Institute of Supervision and Test on Quality & Measurement, Zhanjiang 524094, ChinaZhanjiang Institute of Supervision and Test on Quality & Measurement, Zhanjiang 524094, ChinaCollege of Food Science and Engineering, South China University of Technology, Guangzhou 510641, ChinaThe study aimed to establish the effective extraction methods for Chlamydomonas reinhardtii proteins and screen peptide segments with inhibitory effects on monoamine oxidase A (MAO-A). The proteins were extracted, separated, and purified by swelling, ultrasonication, alkaline dissolution, and salting-out. Subsequently, the polypeptide content, amino acid composition, molecular weight distribution and the MAO-A inhibitory effect of proteins treated with enzymatic hydrolysis and ultrafiltration were determined. Peptide sequences with high inhibitory potential were selected by bioinformatics tools and molecular docking with MAO-A was applied to analyze the binding sites. The results indicated that a protein extraction yield of 50.98%±2.50% was achieved through swelling for 4 h, followed by four cycles of 240 W ultrasonication (10 min per cycle), alkaline dissolution at pH9.5, acid precipitation at pH3.5, and the treatment with saturated ammonium sulfate at a ratio of 1:2. Notably, the alkaline protease hydrolysate (APH) and neutral protease hydrolysate (NPH) exhibited high inhibitory effects on MAO-A (P<0.05), with IC50 values of 3.437 and 2.699 mg/mL, respectively. The fractions with molecular weights less than 3 kDa from APH and NPH (APH-Ⅲ and NPH-Ⅲ) displayed significantly higher MAO-A inhibitory effect (P<0.05), with IC50 values of 3.116 and 0.909 mg/mL, respectively. Molecular docking results showed that two peptides, ASDDAVHGWGGPGGY and AYSGETQSEGGFAPNR from NPH-Ⅲ, could effectively bind to the active sites His488, Glu286, Tyr53, Tyr268, Asp343 and Glu485 of MAO-A.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024070176chlamydomonas reinhardtiiprotein extractionmao-a inhibitory peptidesmolecular docking
spellingShingle Meiting LIU
Ya MAO
Jintao XIE
Wenxia LIU
Lixia WU
Yishan WU
Mei WANG
Xuewu ZHANG
Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory Peptides
Shipin gongye ke-ji
chlamydomonas reinhardtii
protein extraction
mao-a inhibitory peptides
molecular docking
title Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory Peptides
title_full Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory Peptides
title_fullStr Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory Peptides
title_full_unstemmed Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory Peptides
title_short Extraction of Proteins from Chlamydomonas reinhardtii and Screening of MAO-A Inhibitory Peptides
title_sort extraction of proteins from chlamydomonas reinhardtii and screening of mao a inhibitory peptides
topic chlamydomonas reinhardtii
protein extraction
mao-a inhibitory peptides
molecular docking
url http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024070176
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