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Effect of Temperature-Shift and Temperature-Constant Cultivation on the Monacolin K Biosynthetic Gene Cluster Expression in Monascus sp.

ABSTRACT: In this study, the effects of temperature-shift (from 30 to 25 °C) and temperature-constant (at 30 °C) cultivation on the mass of Monascus fuliginosus CG-6 mycelia and concentration of the produced monacolin K (MK) were monitored. The expression levels of the MK biosynthetic genes of M. fuliginosus CG-6 at constant and variable culture temperatures were analysed by real-time quantitative polymerase chain reaction (RT-qPCR). The total protein was collected and determined by liquid chromatography-electrospray ionisation with tandem mass spectrometry (LC-ESI-MS/MS). Results showed that the maximum mycelial mass in temperature-shift cultivation was only 0.477 g of dry cell mass per dish, which was lower than that in temperature-constant cultivation (0.581 g of dry cell mass per dish); however, the maximum concentration of MK in temperature-shift cultivation (34.5 µg/mL) was 16 times higher than that in temperature-constant cultivation at 30 °C (2.11 µg/mL). Gene expression analysis showed that the expression of the MK biosynthetic gene cluster at culture temperature of 25 °C was higher than that at 30 °C, which was similar to the trend of the MK concentration, except for individual MK B and MK C genes. Analysis of differential protein expression revealed that 2016 proteins were detected by LC-ESI-MS/MS. The expression level of efflux pump protein coded by the MK I gene exhibited the same upregulated trend as the expression of MK I in temperature-shift cultivation. Temperature-shift cultivation enhanced the expression of proteins in the secondary metabolite production pathway, but suppressed the expression of proteins involved in the mycelial growth.

SUBMITTER: Lin L

PROVIDER: S-EPMC5434371 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Effect of Temperature-Shift and Temperature-Constant Cultivation on the Monacolin K Biosynthetic Gene Cluster Expression in Monascus sp.

Lin Lin L Wang Changlu C Li Zhenjing Z Wu Huijia H Chen Mianhua M

Food technology and biotechnology 20170301 1

In this study, the effects of temperature-shift (from 30 to 25 °C) and temperature-constant (at 30 °C) cultivation on the mass of Monascus fuliginosus CG-6 mycelia and concentration of the produced monacolin K (MK) were monitored. The expression levels of the MK biosynthetic genes of M. fuliginosus CG-6 at constant and variable culture temperatures were analysed by real-time quantitative polymerase chain reaction (RT-qPCR). The total protein was collected and determined by liquid c ...[more]

PMID: 28559732

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Project description:BackgroundMedical plants confer various benefits to human health and their bioconversion through microbial fermentation can increase efficacy, reduce toxicity, conserve resources and produce new chemical components. In this study, the cholesterol-lowering monacolin K genes and content produced by Monascus species were identified. The high-yield monacolin K strain further fermented with various medicinal plants. The antioxidant and anti-inflammatory activities, red pigment and monacolin K content, total phenolic content, and metabolites in the fermented products were analyzed.ResultsMonacolin K was detected in Monascus pilosus (BCRC 38072), and Monascus ruber (BCRC 31533, 31523, 31534, 31535, and 33323). It responded to the highly homologous mokA and mokE genes encoding polyketide synthase and dehydrogenase. The high-yield monacolin K strain, M. ruber BCRC 31535, was used for fermentation with various medicinal plants. A positive relationship between the antioxidant capacity and total phenol content of the fermented products was observed after 60 days of fermentation, and both declined after 120 days of fermentation. By contrast, red pigment and monacolin K accumulated over time during fermentation, and the highest monacolin K content was observed in the fermentation of Glycyrrhiza uralensis, as confirmed by RT-qPCR. Moreover, Monascus-fermented medicinal plants including Paeonia lactiflora, Alpinia oxyphylla, G. uralensis, and rice were not cytotoxic. Only the product of Monascus-fermented G. uralensis significantly exhibited the anti-inflammatory capacity in a dose-dependent manner in lipopolysaccharide-induced Raw264.7 cells. The metabolites of G. uralensis with and without fermentation (60 days) were compared by LC/MS. 2,3-Dihydroxybenzoic acid, 3,4-dihydroxyphenylglycol, and 3-amino-4-hydroxybenzoate were considered to enhance the antioxidant and anti-inflammatory ability.ConclusionsGiven that highly homologous monacolin K and citrinin genes can be observed in Monascus spp., monacolin K produced by Monascus species without citrinin genes can be detected through the complementary methods of PCR and HPLC. In addition, the optimal fermentation time was important to the acquisition of antioxidants, red pigment and monacolin K. These bioactive substances were significantly affected by medicinal plants over fermentation time. Consequently, Monascus-fermented G. uralensis had a broad spectrum of biological activities.

Dataset Information

Effect of Temperature-Shift and Temperature-Constant Cultivation on the Monacolin K Biosynthetic Gene Cluster Expression in Monascus sp.

Publications

Effect of Temperature-Shift and Temperature-Constant Cultivation on the Monacolin K Biosynthetic Gene Cluster Expression in <i>Monascus</i> sp.

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