Discovery and Engineering of Cytochrome P450s for Terpenoid Biosynthesis

Xiao, Han; Zhang, Yue; Wang, Meng

doi:10.1016/j.tibtech.2018.11.008

Cited by 61 publications

(40 citation statements)

References 79 publications

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“…Although strategies focused on fermentation process optimization of G. lucidum greatly improved GAs content over the past decades, bioproduction of GAs still cannot meet the requirements for the wide range of application (Xu & Zhong, ). Further increasing GAs’ production in G. lucidum fermentation is of particular importance but challenging due to (a) the slow growth of G. lucidum , (b) the notoriously immature genetic manipulations of medicinal mushroom (Xiao & Zhong, ), and (c) the cryptic biosynthetic pathway of GAs (Xiao et al, ).…”

Section: Introductionmentioning

confidence: 99%

Efficient biosynthesis of antitumor ganoderic acid HLDOA using a dual tunable system for optimizing the expression of CYP5150L8 and a Ganoderma P450 reductase

Lan

Wei

Wang

et al. 2019

Biotech & Bioengineering

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Ganoderic acid 3-hydroxy-lanosta-8,24-dien-26-oic acid (GA-HLDOA), an antitumor triterpenoid from the traditional Chinese medicinal higher fungus Ganoderma lucidum, is considered as a key precursor for biosynthesizing other ganoderic acids (GAs) with superior antitumor activities. Our previous study identified CYP5150L8 from G. lucidum as a lanosterol oxidase, and achieved heterologous biosynthesis of GA-HLDOA in Saccharomyces cerevisiae. However, low production of GA-HLDOA in either G. lucidum or heterologous host hindered its further investigation and application. In this study, we constructed a dual tunable system for balancing the expression of CYP5150L8 and a Ganoderma P450 reductase iGLCPR, and performed a comprehensive optimization of CYP5150L8 expression, iGLCPR expression, and glycerol usage. Then, we investigated the fermentation behavior of the best strain in optimized condition in flask and achieved 154.45 mg/L GA-HLDOA production, which was 10.7-fold higher compared with previous report. This study may facilitate the wide-spread application of GA-HLDOA and the discovery of unknown cytochrome P450s in downstream GAs biosynthesis.

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Section: Introductionmentioning

confidence: 99%

Efficient biosynthesis of antitumor ganoderic acid HLDOA using a dual tunable system for optimizing the expression of CYP5150L8 and a Ganoderma P450 reductase

Lan

Wei

Wang

et al. 2019

Biotech & Bioengineering

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“…Among terpenoids, only monoterpenes (C10) were detected in this study, most likely generated via the methylerythritol-4-phosphate (MEP) pathway [28]. In Figure 1A the possible mechanism for the formation of these monoterpenes is given with reference of literatures [29][30][31].…”

Section: Terpenoidsmentioning

confidence: 75%

Free and Bound Aroma Compounds of Turnjujube (Hovenia acerba Lindl.) during Low Temperature Storage

Yang

et al. 2020

Foods

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Free and bound aroma volatiles from turnjujube during low temperature storage were extracted by headspace solid-phase microextraction. They were then characterized and identified using gas chromatography–mass spectrometry. Turnjujube was harvested and stored for 7, 14, and 21 days at 7 °C, the common temperature of display refrigerators in grocery stores. The results showed that 41 free and 24 bound aroma compounds were detected for the first time in turnjujube in both freshly harvested and stored turnjujube. The free and bound aroma compounds of turnjujube were markedly influenced by the storage time. The major free aroma compounds in turnjujube included esters, alcohols, aliphatic aldehydes, and aliphatic ketones. The major bound aroma compounds included borneol, eugenol, and isoeugenol, which contributed to sweet, floral, and herbaceous aroma after their hydrolysis. Freshly harvested turnjujube mostly had a fruity and herbaceous aroma, which diminished after storage at 7 °C. In contrast, the fatty aroma enhanced gradually over storage, and the floral aroma enhanced noticeably after storage for seven days. Foul odor was not detected even after storage at 7 °C for 21 days. The formation mechanisms of some aroma compounds were proposed.

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“…Formation of known cyclic triterpenoids predominately involves cyclization of 2,3-oxidosqualene by terpenoid cyclase, and post-modification by other enzymes including CYPs (Dong et al 2018). Harnessing the great power of protein engineering on terpenoid cyclases and the promiscuity of CYPs (Xiao et al 2019), structurally diversified cyclic triterpenoids can be generated from alternative substrates, including the novel STs discovered in this study.…”

Section: Heterologous Bioproduction Of St-1 St-2 and St-3mentioning

confidence: 91%

“…To circumvent the immature genetic manipulation of mushrooms, we recently developed a synthetic biology platform to identify mushroom-derived CYPs using Saccharomyces cerevisiae as a screening host (Wang et al 2018). We chose S. cerevisiae because: (i) it is a genetically tractable host; (ii) it naturally produces direct triterpenoid precursors (e.g., squalene, lanosterol); (iii) it has subcellular organelles (e.g., endoplasmic reticulum) to support the function of membrane-bounded CYPs; and (iv) its well-characterized endogenous CYPs are less likely to interfere with exogenous ones (Kelly et al 2001;Xiao et al 2019). This strategy enabled the discovery of cyp5150l8 from a famous traditional medicinal mushroom, Ganoderma lucidum, as the first CYP responsible for biosynthesis of the cyclic triterpenoid ganoderic acid HLDOA (Wang et al 2018).…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis of squalene-type triterpenoids in Saccharomyces cerevisiae by expression of CYP505D13 from Ganoderma lucidum

Song

Xiao

Luo

et al. 2019

Bioresour. Bioprocess.

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Background: Squalene-type triterpenoids (STs) are a class of linearized triterpenoids with significant bioactivities, including anti-cancer, anti-oxidative, and anti-inflammatory activities. The efficient biosynthesis of STs has gained increasing attention. Results: Using Saccharomyces cerevisiae as a heterologous host, we discovered that overexpression of CYP505D13 from Ganoderma lucidum, a famous medicinal mushroom capable of producing various triterpenoids as secondary metabolites, enables the engineered S. cerevisiae strain to produce two new STs, 4,8-dihydroxy-22,23-oxidosqualene (ST-1), 8-hydroxy-2,3;22,23-squalene dioxide (ST-2), and a known ST, 2,3; 22,23-squalene dioxide (ST-3), at the respective titers of 3.28 mg/L, 13.77 mg/L, and 12.23 mg/L after 59 h fermentation. Furthermore, our in vitro enzymatic assay implies that CYP505D13 is involved in the formation of ST-3. Conclusions: This study provides a promising alternative to discover STs and facilitate their efficient bioproduction.

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Discovery and Engineering of Cytochrome P450s for Terpenoid Biosynthesis

Cited by 61 publications

References 79 publications

Efficient biosynthesis of antitumor ganoderic acid HLDOA using a dual tunable system for optimizing the expression of CYP5150L8 and a Ganoderma P450 reductase

Efficient biosynthesis of antitumor ganoderic acid HLDOA using a dual tunable system for optimizing the expression of CYP5150L8 and a Ganoderma P450 reductase

Free and Bound Aroma Compounds of Turnjujube (Hovenia acerba Lindl.) during Low Temperature Storage

Biosynthesis of squalene-type triterpenoids in Saccharomyces cerevisiae by expression of CYP505D13 from Ganoderma lucidum

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