Biotechnology for Biofuels
·
2019
·Vol. 12
·期 1
·1-18
·
IF 5.452
Pang Yaru,Zhao Yakun,Li Shenglong,Zhao Yu,Li Jian,Hu Zhihui,Zhang Cuiying,Xiao Dongguang,Yu Aiqun
Background Limonene is an important biologically active natural product widely used in the food, cosmetic, nutraceutical and pharmaceutical industries. However, the low abundance of limonene in plants renders their isolation from plant sources non-economically viable. Therefore, engineering microbes into microbial factories for producing limonene is fast becoming an attractive alternative approach that can overcome the aforementioned bottleneck to meet the needs of industries and make limonene production more sustainable and environmentally friendly. Results In this proof-of-principle study, the oleaginous yeast Yarrowia lipolytica was successfully engineered to produce both d -limonene and l -limonene by introducing the heterologous d -limonene synthase from Citrus limon and l -limonene synthase from Mentha spicata , respectively. However, only 0.124 mg/L d -limonene and 0.126 mg/L l -limonene were produced. To improve the limonene production by the engineered yeast Y. lipolytica strain, ten genes involved in the mevalonate-dependent isoprenoid pathway were overexpressed individually to investigate their effects on limonene titer. Hydroxymethylglutaryl-CoA reductase (HMGR) was found to be the key rate-limiting enzyme in the mevalonate (MVA) pathway for the improving limonene synthesis in Y. lipolytica . Through the overexpression of HMGR gene, the titers of d -limonene and l -limonene were increased to 0.256 mg/L and 0.316 mg/L, respectively. Subsequently, the fermentation conditions were optimized to maximize limonene production by the engineered Y. lipolytica strains from glucose, and the final titers of d -limonene and l -limonene were improved to 2.369 mg/L and 2.471 mg/L, respectively. Furthermore, fed-batch fermentation of the engineered strains Po1g KdHR and Po1g KlHR was used to enhance limonene production in shake flasks and the titers achieved for d -limonene and l -limonene were 11.705 mg/L (0.443 mg/g) and 11.088 mg/L (0.385 mg/g), respectively. Finally, the potential of using waste cooking oil as a carbon source for limonene biosynthesis from the engineered Y. lipolytica strains was investigated. We showed that d -limonene and l -limonene were successfully produced at the respective titers of 2.514 mg/L and 2.723 mg/L under the optimal cultivation condition, where 70% of waste cooking oil was added as the carbon source, representing a 20-fold increase in limonene titer compared to that before strain and fermentation optimization. Conclusions This study represents the first report on the development of a new and efficient process to convert waste cooking oil into d -limonene and l -limonene by exploiting metabolically engineered Y. lipolytica strains for fermentation. The results obtained in this study lay the foundation for more future applications of Y. lipolytica in converting waste cooking oil into various industrially valuable products.
研究领域:
生物技术
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代谢工程
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微生物学
·
可持续化学
·
生物化学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF MEDICAL ENTOMOLOGY
·
2022
·Vol. 59
·期 5
·1669-1677
·
IF 2.435
Jian Rongchao,Lin Yuan,Li Yuling,Wu Weifeng,Ren Xiaofei,Liang Zhanyuan,Kong Lingjia,Cai Jinglin,Lao Canyao,Wu Min,Chen Wenhua,Chen Jing,Hong Weiqian David,Sheng Zhaojun
Aedes albopictus (Skuse) is a vector of several arboviruses, such as dengue, chikungunya, West Nile, and Zika viruses. At present, the use of synthetic insecticides is the main vector control strategy. However, the widespread and long-term use of insecticides has aroused several problems, including insecticide resistance, environmental pollution, and non-target species effects, thereby encouraging researchers to search for new alternatives derived from natural products. In recent decades, essential oils (EOs) as natural alternatives to control mosquitoes have received increasing attention. In the initial larvicidal activity screen, two Rutaceae plants (Citrus aurantium and Citrus paradisi) EOs were selected and evaluated for killing Ae. albopictus larvae. The LC50 values of C. aurantium and C. paradisi EOs against Ae. albopictus were 91.7 and 100.9 ppm, respectively. The main components of C. aurantium EO include diethyl o-phthalate (37.32%), limonene (10.04%), and methyl dihydrojasmonate (6.48%). The main components of C. paradisi EO include limonene (60.51%), diethyl o-phthalate (11.75%), linalool (7.90%), and styralyl acetate (6.28%). Among these main components of the two EOs, limonene showed potent larvicidal activity, with the LC50 value of 39.7 ppm. The nanoemulsions of limonene were prepared and characterized. The duration of larvicidal activity was greater in the limonene nanoemulsions than when limonene was applied in solvent. This study demonstrates that EOs of plants in family Rutaceae are a potential resource to develop new larvicides, and nanoemulsification is an effective method for improving the physicochemical properties and efficacy of natural products as larvicides.
研究领域:
昆虫学
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植物化学
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公共卫生
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
·
2025
·
IF 1.9
Zehao Li,Zixuan Wang,Lulu Zhang,Feng Yingjie,Jinchu Yang,Haoliang Li,Weimin Song,Qiuling Wang,Yongfeng Yang,Zhenzhen Huang,Yongming Xu,Yufeng Fu,Meizhou Ding,Gang Fan,Jingnan Ren,Aiqun Yu,Jian Feng,Guijie Li,Linhua Huang,Yujiao Cheng
α-Terpineol, an abundant oxygenated monoterpene compound, shows a varied range of beneficial bioactivities. This study focused on the efficient extraction and purification of α-terpineol from the biotransformation of limonene using Penicillium digitatum DSM 62840 mutant (PdTP1-overexpressed OE2 strain). The α-terpineol was primarily distributed in the supernatant, with minimal association with the biomass. And the optimal extraction was achieved using ethyl acetate as the extractant, which can directly obtain the highest recovery of α-terpineol without sequential extractions. The equilibrium of mass transfer was quickly reached (≤20 s in vortex). Additionally, the purification method of α-terpineol using column chromatography was described, further improving the purity of product to 96.86% in gas chromatography. Structural identification of purified α-terpineol was confirmed using gas chromatography-mass spectrometry, Fourier-transform infrared spectroscopy, and nuclear magnetic resonance. This simplified and efficient strategy for the extraction and purification of α-terpineol not only provides a solid theoretical basis for the final step in the relevant study of microbial synthesis of α-terpineol but also is of great significance for its industrial application.
研究领域:
微生物学
·
天然产物化学
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药理学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Environmental Sciences
·
2024
·
IF 5.9
Yingqi Zhao,Zhaoyan Zhang,Ya Zhao,Chong Wang,Hua Xie,Jiayue Yang,Weiqing Zhang,Guorong Wu,Gang Li,Ling Jiang,Xueming Yang
It is highly challenging to precisely compare the impacts of anthropogenic pollutants on the photooxidation of isomeric volatile organic compounds with respect to molecular compositions and particle number/mass concentrations of secondary organic aerosols (SOAs). In this study, we conducted a series of well-defined indoor chamber experiments to compare the effects of NO x (NO and NO 2 ) on the photooxidation of isomeric monoterpenes of β -pinene and limonene. For the photooxidation of β -pinene with NO x , the increase of the initial concentrations of NO ([NO] 0 ) shows a monotonous suppression of the particle mass concentration, whereas the increase of [NO 2 ] 0 shows a monotonous enhancement of the particle mass concentration. For the photooxidation of limonene with NO x , the increase of [NO] 0 exhibits a monotonous suppression of the particle mass concentration, whereas the increase of [NO 2 ] 0 shows a parabolic trend of the particle mass concentration. Utilizing a newly developed vacuum ultraviolet free electron laser (VUV-FEL), the online threshold photoionization mass spectrometry reveals a series of novel compounds at molecular weight (MW) = 232 and 306 for the β -pinene + NO x system and MW = 187, 261, 280, and 306 for the limonene + NO x system. The molecular structures and formation pathways of these species were inferred, which led to the prediction of the diversity and difference of SOA products (i.e., ester and peroxide accretion products) formed from different monoterpene precursors. To improve the predictions of future air quality, it is recommended that climate models should incorporate the NO x -driven diurnal photooxidation of monoterpenes for SOA formation mechanisms.
研究领域:
大气化学
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环境科学
✓ 已验证
DOI
高影响因子
FLAVOUR AND FRAGRANCE JOURNAL
·
2022
·Vol. 37
·期 4
·254-261
·
IF 2.5
Zongquan Li,Yunjian Ma,Frank Hollmann,Yonghua Wang
Carvol and carvone are oxidation products from the natural product limonene. They are important raw materials for the flavours and fragrances industry and also act as pharmaceutical active ingredients. Orange waste peels possibly represent an attractive source for limonene, but studies on valorizing orange peel wastes are rare. In this study, we report a new enzymatic cascade system for the in-situ conversion of limonene from orange peel into valued-added carvol and carvone. The use of deep eutectic solvents (DES) allows for efficient in-situ extraction of limonene from waste orange peels. We propose a dual function use of DES as solvent for the extraction and the biocatalytic oxidation of limonene as well as cosubstrate to promote the oxidation reaction. Using ChCl-Pro-H 2 O DES for the extraction of limonene from waste orange peels, approximately 17 milligrams of limonene per gram of orange peel was achieved at 40°C for 24 h. Then, with ChCl-Pro-H 2 O DES as the extractant and reaction medium, a cascade reaction system of choline oxidase (ChOx) and unspecific peroxygenase (UPO) was established to catalyse the conversion of limonene into carvol and carvone. The concentration of the final products was up to about 1.6 mmol L −1 . This study showed a biocatalytic transformation pathway and provides technical support for the high-value utilization of waste in orange peel.
研究领域:
生物化学
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环境科学
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香料工业
✓ 已验证
DOI
高影响因子
JOURNAL OF ECONOMIC ENTOMOLOGY
·
2017
·Vol. 110
·期 4
·1556-1562
·
IF 1.824
Hu Wei,Zhang Ning,Chen Hongli,Zhong Balian,Yang Aixue,Kuang Fan,Ouyang Zhigang,Chun Jiong
Sweet orange oil fractions were prepared by molecular distillation of cold-pressed orange oil from sample A (Citrus sinensis (L.) ‘Hamlin’ from America) and sample B (Citrus sinensis Osbeck ‘Newhall’ from China) respectively, and their fumigant activities against medium workers of red imported fire ants (Solenopsis invicta Buren) were investigated. The volatile composition of the orange oil fractions was identified and quantified using GC–MS. Fractions from sample A (A1, A2, and A3) contained 23, 37, and 48 chemical constituents, and fractions from sample B (B1, B2, and B3) contained 18, 29, and 26 chemical constituents, respectively. Monoterpenes were the most abundant components, accounting for 73.56% to 94.86% of total orange oil fractions, among which D-limonene (65.28–80.18%), β-pinene (1.71–5.58%), 3-carene (0.41–4.01%), β-phellandrene (0.58–2.10%), and linalool (0.31–2.20%) were major constituents. Fumigant bioassay indicated that all orange oil fractions exerted good fumigant toxicity against workers of fire ants at 3, 5, 10, and 20 mg/centrifuge tubes, and B1 had the strongest insecticidal potential, followed by A1, B2, A2, B3, and A3. The fractions composed of more high volatile molecules (A1 and B1) showed greater fumigant effects than others. Compounds linalool and D-limonene, which were the constituents of the orange oil, exhibited excellent fumigant toxicity against red imported fire ant workers. Linalool killed red imported fire ant workers completely at 5, 10, and 20 mg/tube after 8 h of treatment, and D-limonene induced >86% mortality at 8 h of exposure.
研究领域:
昆虫学
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植物化学
✓ 已验证
DOI
PubMed
高影响因子
INDUSTRIAL CROPS AND PRODUCTS
·
2024
·
IF 5.6
Kexin Zhu,Xun Wang,Hao Shi,Fei Wang,Xun Li
Bergamot essential oil has rich biological activity and is commonly applied in fragrance, cosmetics and daily chemical products. However, the entire production process of bergamot fruit from cultivation to extraction of essential oil is inefficient and unstable, cannot meet the growing market demand. To overcome this limitation, we first established a highly efficient biosynthetic platform for production of the major components ((+)-limonene and (-)-linalool) of bergamot essential oil. By increasing the supply of monoterpene precursors and screening for the direct precursors NPP and GPP, a biosynthetic pathway capable of producing both (+)-limonene and (-)-linalool was constructed in Escherichia coli . Then, by screening and mutating the linalool synthase and modifying the limonene synthase by fusion proteins , the obtained strain BW4M42 can produce 277.8 mg/L (+)-limonene and 54.2 mg/L (-)-linalool in shake flasks after optimizing the fermentation culture conditions. In a 1.3-L reactor, the total concentration reached 2688.2 mg/L, including 2010.2 mg/L (+)-limonene and 678 mg/L (-)-linalool. The (-)-linalool/(+)-limonene ratio produced by the strain BW4M42 meets the ISO 3520:2022 standard for bergamot essential oil. This study provides the possibility to construct a biosynthetic platform for producing the major components of plant essential oils through metabolic engineering .
研究领域:
生物化学
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合成生物学
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微生物学
✓ 已验证
DOI
高影响因子
Foods
·
2025
·Vol. 14
·期 9
·1637
·
IF 5.1
Weiheng Kong,Xianjiang Lin,Jing Ye,Yanbin Lu
In this study, an efficient and eco-friendly method was developed for continuous liquid–liquid extraction of lutein from microalgaeChlorella vulgaris. By employing a limonene-based biphasic liquid system, high-purity lutein was successfully obtained from the crude extract in a single run via centrifugal partition chromatography (CPC). Evaluation and optimization results demonstrated that limonene could effectively serve as a replacement forn-hexane as the solvent system for lutein extraction, exhibiting natural renewability and minimal environmental impact. Furthermore, the elution–extrusion operation mode was employed to fully exploit the liquid nature of the stationary phase in the extraction process, allowing for continuous sampling and separation without interruption. This proposed protocol offers a sustainable and environmentally friendly alternative for extracting valuable ingredients from microalgae biomass, demonstrating its potential as a scalable solution for producing lutein-enriched ingredients applicable to functional foods and nutraceuticals.
研究领域:
生物技术
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环境科学
·
食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2021
·Vol. 28
·期 42
·59665-59672
·
IF 4.223
Zhang Ning,Liao Yihong,Xie Lianjie,Zhang Zhixiang,Hu Wei
The red imported fire ant is one of the world’s most devastating invasive species, adversely affecting humans, wildlife, crops, and livestock. To control infestations, chemical pesticides are deployed extensively around the world. However, their extensive use has led to negative effects on the environment and human health. Essential oils, which are safe and ecofriendly, can potentially be used as alternatives to chemical pesticides. In this study, grapefruit essential oils were used as fumigant agents to control red imported fire ants. The crude grapefruit oil (GO1) contained 28 compounds, and the concentrated grapefruit oil (GO2), which was refined from GO1 by vacuum distillation, contained 20 compounds. D-Limonene was the dominant constituent in both GO1 (70.1%) and GO2 (73.96%), and other important constituents included β-pinene, α-pinene, β-phellandrene, octanal, d-carvone, α-terpineol, and linalool. Both the essential oils and their individual constituents (α-pinene, α-terpineol, β-phellandrene, octanal, and d-carvone) showed strong lethal fumigant effects against workers. Workers were more susceptible to GO2 than GO1, and octanal was more toxic to workers as compared with the other four constituents. When antennas of workers were treated with the two oils or the five constituents, their walking and gripping abilities were significantly suppressed, and there was an obvious bending or breaking phenomenon on the sensilla of the antennas. Fumigant activity by grapefruit essential oils and their main compounds were associated with their effects on the walking and gripping behavior of workers, and this confirmed that grapefruit essential oil is a promising, ecofriendly, and safe fumigant for the control of red imported fire ants.
研究领域:
生态学
·
昆虫学
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环境科学
✓ 已验证
DOI
PubMed
高影响因子
Nutrients
·
2025
·Vol. 17
·期 7
·1255
·
IF 5
Jingxin Chen,Genghua Ou,Wenting Gu,Jian Shi,Ruiying Lyu,Xueping Wu,Junming Wang,Chunhong Liu
Background/Objectives: Chronic alcohol overconsumption triggers alcohol liver injury, and therapeutic strategies targeting alcohol-triggered oxidative stress and hepatic inflammatory responses represent potential approaches to ameliorating alcohol-related hepatotoxicity. This study aimed to determine the hepatoprotective activity of finger citron essential oil (FCEO) in alcoholic liver disease (ALD)-afflicted rats and explore its underlying mechanisms. In order to identify the effective components, we compared the effects of FCEO and D-limonene.Methods: The regulatory effects of FCEO on metabolic enzymes were systematically evaluated through in vitro experiments. In vivo studies were conducted to investigate and compare the hepatoprotective effects of FCEO and D-limonene. Staining methods, assay kits, and Western Blot were used to determine the roles of FCEO and D-limonene in the ALD rats.Results: We found that FCEO downregulated phase I metabolic enzymes and upregulated phase II metabolic enzymes in Buffalo Rat Liver-3A (BRL-3A) cells. FCEO and/or D-limonene intervention reduced transaminase levels in ALD rats and effectively alleviated inflammatory cell infiltration and lipid droplet accumulation in their liver tissue. Additionally, FCEO and D-limonene played a regulatory role in oxidative stress and inflammation-related pathways such as the MAPK/Nrf2 and NF-κB/AMPK pathways. FCEO was superior to D-limonene as an antioxidant in alleviating alcoholic liver injury.Conclusions: This study revealed the alleviative effects and mechanisms of FCEO on alcoholic liver injury, demonstrating better efficacy compared to its monomer, thus providing a strategy for the development and utilization of finger citron resources.
研究领域:
药理学
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肝脏病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Insects
·
2024
·Vol. 15
·期 5
·345
·
IF 2.7
Jifan Cao,Wende Zheng,Baizhong Chen,Zhenping Yan,Xiaowen Tang,Jiahao Li,Zhen Zhang,Song Ang,Chen Li,Rihui Wu,Panpan Wu,Wen-Hua Chen
Simple SummaryThe essential oil (EO) ofCitrus reticulataBlanco cv. Chachiensis (Chachi) exhibited significant larvicidal activity. The EO from semi-mature fruits and its main components were able to control mosquitoes by effectively inhibiting acetylcholinesterase and three detoxification enzymes and significantly altering the diversity of the internal microbiota in mosquitoes.AbstractThe overuse of synthetic insecticides has led to various negative consequences, including insecticide resistance, environmental pollution, and harm to public health. This may be ameliorated by using insecticides derived from botanical sources. The primary objective of this study was to evaluate the anti-mosquito activity of the essential oil (EO) ofCitrus reticulataBlanco cv. Chachiensis (Chachi) (referred to as CRB) at immature, semi-mature, and mature stages. The chemical compositions of the CRB EO were analyzed using GC-MS. The main components were identified to be D-limonene andγ-terpinene. The contents of D-limonene at the immature, semi-mature, and mature stages were 62.35%, 76.72%, and 73.15%, respectively; the corresponding contents ofγ-terpinene were 14.26%, 11.04%, and 11.27%, respectively. In addition, the corresponding contents of a characteristic component, methyl 2-aminobenzoate, were 4.95%, 1.93%, and 2.15%, respectively. CRB EO exhibited significant larvicidal activity againstAedes albopictus(Ae. albopictus, Diptera: Culicidae), with the 50% lethal doses being 65.32, 61.47, and 65.91 mg/L for immature, semi-mature, and mature CRB EO, respectively. CRB EO was able to inhibit acetylcholinesterase and three detoxification enzymes, significantly reduce the diversity of internal microbiota in mosquitoes, and decrease the relative abundance of core species within the microbiota. The present results may provide novel insights into the utilization of plant-derived essential oils in anti-mosquitoes.
研究领域:
植物学
·
公共卫生
·
昆虫学
✓ 已验证
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2014
·
IF 3.107
Mohamed Reda Zahi,Pingyu Wan,Hao Liang,Qipeng Yuan
d-Limonene organogel-based nanoemulsion was prepared by high-pressure homogenization technology. The organogelator type had a major role on the formation of the formulations, in which stearic acid has given nanoemulsions with the smallest droplet size. The surfactant type and concentration also had an appreciable effect on droplet formation, with Tween 80 giving a mean droplet diameter (d ≈ 112 nm) among a range of non-ionic surfactants (Tween 20, 40, 60, 80, and 85). In addition, high-pressure homogenization conditions played a key role in the nanoemulsion preparation. The stability of d-limonene organogel-based nanoemulsion was also investigated under two different temperatures (4 and 28 °C) through 2 weeks of storage. Results showed a good stability of the formulations, which is maybe due to the incorporation of d-limonene into the organogel prior to homogenization. This study may have a valuable contribution for the design and use of organogel-based nanoemulsion as a delivery system in food.
研究领域:
食品科学
·
化学工程
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）