|
|
|
| Untargeted Metabolomics Analysis of Products from Paecilomyces cicadae Fermentation of Camellia oleifera Oil Deodorizer Distillate |
| ZHENG Jun-Rong1, CHEN Xiu-Ming1, LE Zhan-Xian1,*, ZHOU A-Rong1, JIA Wei1, WU Long-Jing2, WU Min2, YE Zhu-Fu2, CHEN Jian-Long3, LUO Li-Jin1,* |
1 Fujian Institute of Microbiology, Fuzhou 350007, China; 2 Fujian Chuanjiu Agricultural Development Co., Ltd., Ningde 355502, China; 3 Fuzhou Lianfu Biotechnology Co., Ltd., Fuzhou 350307, China |
|
|
|
|
Abstract Camellia oleifera oil deodorizer distillate (CODD) is rich in high-value lipid components such as phytosterols, vitamin E, and unsaturated fatty acids, yet its direct utilization as a feed additive is hampered by poor dispersibility and low bioavailability. To develop green feed additives as alternatives to dietary antibiotics and achieve high-value utilization of CODD. In this study, Paecilomyces cicadae was inoculated into a solid-state fermentation system with soybean meal and wheat bran as carriers to ferment CODD. Non-targeted metabolomics combined with multivariate statistical analysis, KEGG pathway enrichment, and FELLA(functional enrichment analysis and visualization using sparse network-based activity diffusion) network analysis was employed to systematically elucidate the differential characteristics and regulatory mechanisms of lipid metabolism before and after fermentation. The results showed that the metabolic profiles of the fermentation and control groups exhibited significant separation. The expression patterns of differential lipid metabolites could be categorized into 4 functional groups: Comprehensive up-regulation of flavonoids (e.g., chrysin increased by over 4-fold), bidirectional regulation of steroids (anti-inflammatory pregnanes up-regulated while inert sterones down-regulated), uniform down-regulation of pro-inflammatory prostaglandins, and significant up-regulation of terpenoids. Mechanistic studies revealed that cutin/suberine/wax biosynthesis, steroid hormone biosynthesis, and arachidonic acid metabolism constitute 3 core regulatory pathways. The FELLA (functional enrichment analysis and visualization using sparse network-based activity diffusion) network further elucidated a cross-pathway synergistic regulatory mechanism with diacylglycerol and phosphatidylethanolamine as hub nodes and lipolytic enzymes together with acyltransferases as key enzyme targets. In conclusion, P. cicadae fermentation achieves pre-conversion of inert plant sterols present in CODD into highly active C21 steroids and directional restructuring of hesperetin-type precursor flavonoids into anti-inflammatory flavonoids such as chrysin, thereby enhancing the anti-inflammatory and antioxidant functional potential of the fermented products. This study provides a theoretical basis for the resource utilization of CODD and the development of green feed additives.
|
|
Received: 27 March 2026
|
|
|
|
Corresponding Authors:
*pharmfil@163.com; luolijin@sina.com
|
|
|
|
[1] 陈官菊, 柴一秋, 厉晓腊, 等. 2019. 蝉拟青霉类枯草杆菌蛋白酶基因的克隆及其序列和蛋白质分析[J]. 浙江农业学报, 31(10): 1663-1670. (Chen G J, Chai Y Q, Li X L, et al.2019. Cloning of sublitisin-like protease gene cDNA from Isaria cicadae and characterization of its sequence and deduced protein[J]. Acta Agriculturae Zhejiangensis, 31(10): 1663-1670.) [2] 付建平, 韩晓丹, 胡居吾, 等. 2020. 分子蒸馏技术提取茶油脱臭馏出物中维生素E[J]. 食品工业, 41(9): 186-189. (Fu J P, Han X D, Hu J W, et al.2020. Extraction of vitamin E from camellia oil deodorizer distillate by molecular distillation technology[J]. The Food Industry, 41(9): 186-189.) [3] 刘静洁, 马衍旋, 蒋春玲, 等. 2021. 发酵豆粕的营养价值及其在动物饲料中应用的研究进展[J]. 黑龙江畜牧兽医, (13): 27-30. (Liu J J, Ma Y X, Jiang C L, et al. 2021. Research progress on the nutritional value of fermented soybean meal and its application in animal feed[J]. Heilongjiang Animal Science and Veterinary Medicine, (13): 27-30.) [4] 马新燕, 李大刚, 余苗, 等. 2024. 饲用抗生素替代品在畜禽生产中应用研究进展[J]. 动物营养学报, 36(12): 7502-7512. (Ma X Y, Li D G, Yu M, et al.2024. Research progress on application of feed antibiotic substitutes in livestock and poultry production[J]. Chinese Journal of Animal Nutrition, 36(12): 7502-7512.) [5] 肖希贤. 2022. 茶油脱臭馏出物中甾醇的提取及其对VC脂质体性能的影响[D]. 硕士学位论文, 华南农业大学, 导师: 吴雪辉, pp. 75-89. (Xiao X X.2022. Extraction of sterols from deodorized distillate of camellia oil and its effect on the properties of VC liposomes[D]. Thesis for M.S., South China Agricultural University, Supervisor: Wu X H, pp. 75-89.) [6] 谢娜, 程俊文, 徐娟, 等. 2024. 酶法辅助提取人工培育蝉花多糖工艺优化及其动力学、热力学、抗氧化研究[J]. 食品工业科技, 45(4): 151-160. (Xie N, Cheng J W, Xu J, et al.2024. Process optimization of enzyme-assisted extraction of polysaccharides from artificially cultivated Cordyceps cicadae and its kinetics, thermodynamics and antioxidant research[J]. Science and Technology of Food Industry, 45(4): 151-160.) [7] 熊亮斌, 宋璐, 赵云秋, 等. 2021. 甾体化合物绿色生物制造: 从生物转化到微生物从头合成[J]. 合成生物学, 2(6): 942-963. (Xiong L B, Song L, Zhao Y Q, et al.2021. Green biomanufacturing of steroid compounds: From biotransformation to microbial de novo synthesis[J]. Synthetic Biology, 2(6): 942-963.) [8] 章娜, 杨凯丽, 张沙, 等. 2023. 植物甾醇酯对产蛋末期蛋鸡产蛋性能、蛋品质、肝脏抗氧化能力及卵黄前体物质合成的影响[J]. 动物营养学报, 35(11): 7123-7137. (Zhang N, Yang K L, Zhang S, et al.2023. Effects of phytosterol esters on laying performance, egg quality, liver antioxidant capacity and yolk precursor synthesis of laying hens at late laying stage[J]. Chinese Journal of Animal Nutrition, 35(11): 7123-7137.) [9] Andersen M R, Nielsen M L, Nielsen J, et al.2011. Metabolic model integration of the bibliome, genome, metabolome and reactome of Aspergillus niger[J]. Molecular Systems Biology, 6(1): 431. [10] Bampidis V, Azimonti G, Bastos M, et al.2023. Assessment of the feed additive consisting of Enterococcus lactis DSM 7134 and Lacticaseibacillus rhamnosus DSM 7133 (provita LE) for calves for rearing for the renewal of its authorisation (lactosan GmbH & Co.KG)[J]. EFSA Journal, 21(10): 8350. [11] Bhatt D K, Mehta A A.2014. Emerging role of PGE2 in inflammation, cancer and developmental biology[J]. Journal of Clinical and Diagnostic Research, 8(8): ME01-ME04. [12] Cai S, Li Q, Zhou H, et al.2021. Mechanism of PI3K/AKT/mTOR signaling pathway for mediating anti-inflammatory and anti-oxidant effects of chrysin: A protein microarray-based study[J]. Journal of Southern Medical University, 41(10): 1554-1561. [13] Cortada-Garcia J, Daly R, Arnold S A, et al.2023. Streamlined identification of strain engineering targets for bioprocess improvement using metabolic pathway enrichment analysis[J]. Scientific Reports, 13(1): 12990. [14] Ge C, Li Y C, Gao S J, et al.2018. Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms[J]. Applied Microbiology and Biotechnology, 102(22): 9597-9608. [15] Geistlinger L, Csaba G, Küffner R, et al.2011. From sets to graphs: Towards a realistic enrichment analysis of transcriptomic systems[J]. Bioinformatics, 27(13): i366-i373. [16] Ghannoum M A.2000. Potential role of phospholipases in virulence and fungal pathogenesis[J]. Clinical Microbiology Reviews, 13(1): 122-143. [17] Han C, Shi C, Liu L, et al.2024. Majorbio Cloud 2024: Update single‐cell and multiomics workflows[J]. iMeta,3(4): 1. [18] Heux S, Sablayrolles J M, Cachon R, et al.2006. Engineering a Saccharomyces cerevisiae strain able to grow on cellobiose and to tolerate lignocellulose hydrolysates[J]. Process Biochemistry, 41(12): 2472-2479. [19] Kalaiselvan P, Devi N C, Deepti M, et al.2025. Solid-state fermentation-a sustainable future technology in aquafeeds?[J]. Frontiers in Marine Science, 12: 1669719. [20] Goto S., Hattori M, Itoh M.2008. KEGG for linking genomes to life and the environment[J]. Nucleic Acids Research, 36(1): D480-D484. [21] Kelly S L, Lamb D C, Kelly D E.2018. Cytochrome P450 (CYP) plasticity in the face of a changing world: Insights from comparative genomics[J]. Natural Product Reports, 35(12): 1253-1266. [22] Köhler G A, Namork E, Snellingen M, et al.2006. Phospholipase A2 and phospholipase B activities in fungi[J]. Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids, 1761(11): 1399-1405. [23] Kishimoto S, Hatano A, Kami S, et al.2019. Detecting and dissecting signaling crosstalk via the multilayer network integration of signaling and regulatory interactions[J]. Nature Communications, 10(1): 5378. [24] Kurkiewicz M, Moździerz A, Rzepecka-Stojko A, et al.2025. Chrysin: A comprehensive review of its pharmacological properties and therapeutic potential[J]. Pharmaceuticals, 18(8): 1162. [25] Lee F C, Ahumada G G, Gross R W, et al.2002. Hydrolysis of phosphatidylethanolamine induced by nominally synthetic lysophosphoglycerides: Methodological implications[J]. Biochemistry, 19(9): 1934-1937. [26] Liu Y, Song X, He J, et al.2019. Biotransformation of chrysin by filamentous fungi: Production of chrysin 7-O-β-D-glucopyranoside and chrysin 7-O-β-D-(4'-O-methyl)-glucopyranoside[J]. Molecules, 24(12): 2284. [27] Medina F, Diaz C, Karrow N, et al.2022. Filamentous fungi as cell factories: Molecular insights into substrate sensing and metabolic remodeling[J]. Microbial Biotechnology, 15(3): 856-874. [28] Murakami C, Atsuta‐Tsunoda K, Inomata S, et al.2025. Human PHOSPHO1 exhibits phosphatidylcholine-and phosphatidylethanolamine-phospholipase C activities and interacts with diacylglycerol kinase δ[J]. FEBS Letters, 599(8): 1169-1186. [29] Murakami C, Sakane F.2021. Sphingomyelin synthase-related protein generates diacylglycerol via hydrolysis of phosphatidic acid, phosphatidylcholine, phosphatidylinositol and phosphatidylethanolamine without ceramide[J]. Journal of Biological Chemistry, 296: 100347. [30] Obianwuna U E, Oleforuh-Okoleh V U, Wang J, et al.2022.Potential implications of natural antioxidants of plant origin on oxidative stability of chicken albumen during storage: A Review[J]. Antioxidants (Basel, Switzerland),11(4): 630. [31] Ogawa J, Shimizu S.1999. Microbial enzymes: new industrial applications from traditional screening methods[J]. Trends in Microbiology, 7(7): 282-287. [32] Olivera E R,Luengo J M.2019. Steroids as environmental compounds recalcitrant to degradation: Genetic mechanisms of bacterial biodegradation pathways[J]. Genes, 10(7): 512. [33] Parks D H, Hugenholtz P, Tyson G W, et al.2023. Carbon acquisition ecological strategies to connect soil microbial biodiversity and carbon cycling[J]. Soil Biology and Biochemistry, 177: 108893. [34] Sakane F, Imai S, Kai M, et al.2007. Diacylglycerol kinases: Why so many of them?[J]. Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids, 1771(7): 793-806. [35] Thevenieau F, Nicaud J M, Gaillardin C.2010. Soluble and cell-bound lipases of the yeast Yarrowia lipolytica[J]. Lipid Technology, 22(5): 107-110. [36] Walle T, Otake Y, Brubaker J A, et al.2001. Disposition and metabolism of the flavonoid chrysin in normal volunteers[J]. British Journal of Clinical Pharmacology, 51(2): 143-146. [37] Xu Y, He J, Song X, et al.2023. Metabolism of chrysin by Rhodotorula glutinis: From C-8 hydroxylation to A-ring cleavage[J]. Molecules, 28(9): 3856. |
|
|
|