Kubayi Sagwadi, Makola Raymond Tshepiso, Dithebe Khumiso
Department of Biochemistry, Microbiology and Biotechnology, University of Limpopo, Private Bag X 1106, Sovenga, Polokwane 0727, South Africa.
Microorganisms. 2025 Mar 19;13(3):692. doi: 10.3390/microorganisms13030692.
Fungal endophytes of medicinal plants produce diverse secondary metabolites and extracellular enzymes with therapeutic and biotechnological potential. However, the biological and biotechnological potential of fungal endophytes from South African medicinal plants remain relatively underexplored. In this study, the antimicrobial, antioxidant, anti-inflammatory and extracellular enzymatic capabilities of five fungal endophytes previously isolated from the leaves of Oliv. were investigated. Sequencing of the internal transcribed spacer (ITS) regions revealed that the isolates belonged to the genera , and . The broth micro-dilution assay and the Folin-Ciocalteau reagent method were used to assess the antibacterial activity and the total phenolic content (TPC) of the fungal endophytes' ethyl acetate crude extracts (CEs), respectively. The antioxidant activity was assessed using the ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assays. The influence of the CE of the KaL-4 on the viability and LPS-induced interleukin-6 (IL-6) production in Raw 264.7 macrophages was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and an ELISA, respectively. The ability of the isolates to produce extracellular proteases, laccases and peroxidases was also determined. The CEs displayed antimicrobial activity with MICs ranging from 0.63 to 1.25 mg/mL and reducing power and scavenging activity ranging from 40% to 18% and from 60% to 48%, respectively. The KaL-4 CE possessed the highest TPC and demonstrated dose-dependent cytotoxicity. The CE further demonstrated a significant reduction in IL-6 production at a concentration of 0.75 µg/mL. Only one isolate demonstrated the ability to produce proteases with an enzymatic index (EI) of 0.66, while laccases (EI range of 0.14 to 1.15) and peroxidases were produced by all of the isolates. These findings suggest that fungal endophytes from South African medicinal plants are promising sources of bioactive compounds and industry-significant extracellular enzymes.
药用植物的真菌内生菌可产生具有治疗和生物技术潜力的多种次生代谢产物和细胞外酶。然而,南非药用植物真菌内生菌的生物学和生物技术潜力仍相对未得到充分探索。在本研究中,对先前从Oliv.叶片中分离出的五种真菌内生菌的抗菌、抗氧化、抗炎和细胞外酶活性进行了研究。内部转录间隔区(ITS)区域测序显示,分离株属于、和属。分别采用肉汤微量稀释法和福林-酚试剂法评估真菌内生菌乙酸乙酯粗提物(CEs)的抗菌活性和总酚含量(TPC)。采用铁还原抗氧化能力(FRAP)和1,1-二苯基-2-苦基肼(DPPH)自由基清除试验评估抗氧化活性。分别采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)试验和酶联免疫吸附测定(ELISA)评估KaL-4的CE对Raw 264.7巨噬细胞活力和脂多糖诱导的白细胞介素-6(IL-6)产生的影响。还测定了分离株产生细胞外蛋白酶、漆酶和过氧化物酶的能力。CEs表现出抗菌活性,最低抑菌浓度(MICs)范围为0.63至1.25 mg/mL,还原能力和清除活性分别为40%至18%和60%至48%。KaL-4的CE具有最高的TPC,并表现出剂量依赖性细胞毒性。该CE在浓度为0.75 µg/mL时还显著降低了IL-6的产生。只有一株分离株表现出产生蛋白酶的能力(酶指数(EI)为0.66),而所有分离株均产生漆酶(EI范围为0.14至1.15)和过氧化物酶。这些发现表明,南非药用植物的真菌内生菌是生物活性化合物和具有工业意义的细胞外酶的有前途的来源。