Feasible biosynthesis of biologically active metabolites in in vitro culture of Macrotyloma uniflorum

IF 3.4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biocatalysis and agricultural biotechnology Pub Date : 2024-10-01 DOI:10.1016/j.bcab.2024.103374

Sannia Batool , Mehnaz Khanum , Hasnat Tariq , Gouhar Zaman , Muhammad Uzair Javed , Bilal Haider Abbasi , Christophe Hano , Nathalie Giglioli-Guivarc'h

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引用次数: 0

Abstract

Macrotyloma uniflorum, commonly called “Horse gram” is an underutilized pulse crop recognized for its great nutritional significance and a broad range of biological properties. There have been no in vitro studies for the biosynthesis of enhanced bioactive metabolites in the callus culture of M. uniflorum. In the study reported here, we have designed a feasible protocol for high-frequency callus induction and maintenance utilizing a leaf as an explant grown on MS media enriched with various concentrations of different plant growth regulators (PGRs) including α-naphthalene acetic acid (NAA), 6-benzylaminopurine (BAP) and thidiazuron (TDZ) either alone or in combination. Among all the tested PGRs, NAA alone resulted in high-frequency callus induction (97%), and maximum biomass accumulation (Fresh weight (FW): 200 g/L: Dry weight (DW): 20.4 g/L). Moreover, hormonally optimized callus cultures exhibit maximum production of phenolic compounds (166.6 mg/L) and flavonoid compounds (351.6 mg/L). The antioxidant potential of calli extracts was also determined by utilizing various antioxidant activities. Maximum antioxidant activities ((2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)) ABTS = 497.9; TEAC μM; (Ferric reducing antioxidant power) FRAP = 823 TEAC μM; (Cellular Antioxidant Activity) CAA: 58.2%) were recorded in leaf-derived calli supplemented with different PGRs treatments. High-performance liquid chromatography (HPLC) analysis further revealed maximum biosynthesis of caffeic acid (1.63 mg/g DW), gallic acid (8.92 mg/g DW), kaempferol (0.71 mg/g DW), myricetin (0.39 mg/g DW), apigenin (0.64 mg/g DW) at 10 mg/L BAP and isorhamnetin (0.68 mg/g DW) at 1 mg/L TDZ + 10 mg/L NAA. The objective of this study was to explore in vitro biosynthesis of enhanced bioactive metabolites in the callus culture of M. uniflorum, leveraging a feasible protocol for high-frequency callus induction and maintenance. The results showcase the remarkable efficacy of NAA in callus induction and biomass accumulation, highlighting the hormonally optimized callus cultures as a potent source for enhanced biosynthesis of bioactive metabolites, paving the way for further applications in pharmaceutical and commercial industries.

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在巨藻体外培养中生物活性代谢物的可行生物合成

大花荠属（Macrotyloma uniflorum），俗称 "马齿苋"，是一种未充分利用的豆类作物，因其重要的营养价值和广泛的生物特性而广受认可。目前还没有关于在马齿苋胼胝体培养中生物合成增强生物活性代谢物的体外研究。在本文报告的研究中，我们设计了一种利用叶片作为外植体在富含不同浓度植物生长调节剂（PGRs）（包括α-萘乙酸（NAA）、6-苄基氨基嘌呤（BAP）和噻虫嗪（TDZ））的 MS 培养基上生长的高频率胼胝体诱导和维持的可行方案，PGRs 包括α-萘乙酸（NAA）、6-苄基氨基嘌呤（BAP）和噻虫嗪（TDZ）。在所有测试的 PGRs 中，单用 NAA 可诱导出高频率的胼胝体（97%）和最大生物量积累（鲜重 (FW)：200 克/升：干重（DW）：20.4 克/升）。此外，激素优化的胼胝体培养物显示出最大的酚类化合物产量（166.6 毫克/升）和黄酮类化合物产量（351.6 毫克/升）。还利用各种抗氧化活性测定了胼胝体提取物的抗氧化潜力。在补充了不同 PGRs 处理的叶生胼胝体中记录到了最高的抗氧化活性（（2,2′-偶氮-双（3-乙基苯并噻唑啉-6-磺酸））ABTS = 497.9；TEAC μM；（铁还原抗氧化能力）FRAP = 823 TEAC μM；（细胞抗氧化活性）CAA：58.2%）。高效液相色谱（HPLC）分析进一步显示，在 10 mg/L BAP 和 1 mg/L TDZ + 10 mg/L NAA 条件下，咖啡酸（1.63 mg/g DW）、没食子酸（8.92 mg/g DW）、山柰酚（0.71 mg/g DW）、杨梅素（0.39 mg/g DW）、芹菜素（0.64 mg/g DW）和异鼠李素（0.68 mg/g DW）的生物合成量最大。本研究的目的是利用高频率诱导和维持胼胝体的可行方案，探索增强生物活性代谢物在单叶芸香科植物胼胝体培养中的体外生物合成。结果表明，NAA 在胼胝体诱导和生物量积累方面具有显著功效，突出表明激素优化的胼胝体培养物是增强生物活性代谢物生物合成的有效来源，为进一步应用于制药和商业行业铺平了道路。

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来源期刊

Biocatalysis and agricultural biotechnology Agricultural and Biological Sciences-Agronomy and Crop Science

CiteScore

7.70

自引率

2.50%

发文量

308

审稿时长

48 days

期刊介绍： Biocatalysis and Agricultural Biotechnology is the official journal of the International Society of Biocatalysis and Agricultural Biotechnology (ISBAB). The journal publishes high quality articles especially in the science and technology of biocatalysis, bioprocesses, agricultural biotechnology, biomedical biotechnology, and, if appropriate, from other related areas of biotechnology. The journal will publish peer-reviewed basic and applied research papers, authoritative reviews, and feature articles. The scope of the journal encompasses the research, industrial, and commercial aspects of biotechnology, including the areas of: biocatalysis; bioprocesses; food and agriculture; genetic engineering; molecular biology; healthcare and pharmaceuticals; biofuels; genomics; nanotechnology; environment and biodiversity; and bioremediation.