Sains Malaysiana 54(5)(2025): 1231-1238

http://doi.org/10.17576/jsm-2025-5405-03

Lipid Metabolon in Non-Oleaginous Fungus Aspergillus niger

(Metabolon Lipid dalam Kulat Bukan Oleginous Aspergillus niger)

IZYANTI IBRAHIM¹, SHUWAHIDA SHUIB², YAP YIN XIN¹ & AIDIL ABDUL HAMID^1,*

¹Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

²Allergy and Immunology Research Centre, Institute for Medical Research (IMR), National Institutes of Health (NIH), Jalan Setia Murni U13/52, Seksyen U13 Setia Alam, 40170 Shah Alam, Selangor, Malaysia

Received: 9 November 2023/Accepted: 7 January 2025

Abstract

The lipogenic multienzyme complex or lipid metabolon consisting of malic enzyme (ME), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC) and ATP:citrate lyase (ACL) was observed during lipid biosynthesis of an oleaginous fungus Cunninghamella bainieri 2A1. It is believed that high production of fatty acid in oleaginous species is due to the association of lipogenic enzymes forming a metabolon during lipid accumulation phase. One of the significances of lipid metabolon is that it provides efficient and rapid channel of NADPH directly from ME to FAS when lipid accumulation occurs. This leads to the assumption of low lipid yield in non-oleaginous microorganisms is due to the absence of lipid metabolon in these microorganisms. Thus, the non-oleaginous filamentous fungi Aspergillus niger was studied to see the association of lipid metabolon and lipid biosynthesis. The cells of A. nigerduring both lipid accumulation and lipid cessation phase were used to prepare cell-free extracts using protoplasting technique. The crude proteins were then separated by Blue Native (BN)-PAGE, associated with liquid chromatography-tandem mass spectrometry (LC-MS/MS). No high molecular mass complex was detected, speculating that no lipogenic multienzyme complex was formed in non-oleaginous fungi hence very low or no storage lipid was accumulated in this fungus.

Keywords: Aspergillus niger; lipid metabolon; lipogenesis; non-oleaginous

Abstrak

Kompleks multienzim lipogenik atau metabolon lipid yang terdiri daripada enzim malik (ME), asid lemak sintase (FAS), karboksilase asetil-KoA (ACC) dan ATP: sitrat liase (ACL) didapati terbentuk semasa biosintesis lipid kulat oleaginus Cunninghamella bainieri 2A1. Penghasilan asid lemak jumlah yang tinggi dalam spesies oleaginus dipercayai disebabkan oleh gabungan enzim lipogenik membentuk metabolon semasa fasa pengumpulan lipid. Salah satu kepentingan metabolon lipid adalah untuk membolehkan penyaluran NADPH secara berterusan dari ME ke FAS dengan cekap dan pantas semasa tempoh pengumpulan lipid. Ini membawa kepada andaian bahawa pengumpulan lipid yang rendah dalam mikroorganisma bukan oleaginus adalah disebabkan oleh ketiadaan metabolon lipid dalam mikroorganisma ini. Oleh itu, kulat berfilamen bukan oleaginus Aspergillus niger telah dikaji untuk melihat perkaitan metabolon lipid dan biosintesis lipid. Kultur sel A. niger semasa fasa pengumpulan lipid dan fasa pemberhentian lipid digunakan untuk menyediakan ekstrak bebas sel menggunakan teknik protoplasting. Ekstrak bebas sel kemudiannya dipisahkan menggunakan Blue Native (BN)-PAGE dan dianalisis menggunakan spektrometri jisim tandem kromatografi cecair (LC-MS/MS). Tiada kompleks berjisim molekul besar dikesan, mencadangkan bahawa tiada kompleks multienzim lipogenik terbentuk dalam kulat bukan oleaginus dan seterusnya membawa kepada pengumpulan jumlah lipid yang sangat rendah dalam kulat ini.

Kata kunci: Aspergillus niger; lipogenesis; metabolon lipid; non-oleaginus

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*Corresponding author; email: aidilah@ukm.edu.my