High-performance liquid chromatography/mass spectrometry (HPLC/MS) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used to characterize the procyanidin composition of the grape seed extract. The detection of the oligomers composed of (+)-catechin, (-)-epicatechin, and their galloylated derivatives in the grape seeds is demonstrated. With MALDI-TOF MS, oligomers up to nonamers were observed. The potential of the MALDI-TOF MS technique as a quantification tool is also discussed. The information presented in this study could lead to the determination of procyanidin content and their molecular weight distribution in grape seeds.
The carotenoids produced by extremely halophilic archaeon Haloarcula japonica were extracted and identified by their chemical, chromatographic, and spectroscopic characteristics (UV-Vis and mass spectrometry). The composition (mol%) was 68.1% bacterioruberin, 22.5% monoanhydrobacterioruberin, 9.3% bisanhydrobacterioruberin, <0.1% isopentenyldehydrorhodopin, and trace amounts of lycopene and phytoene. The in vitro scavenging capacity of a carotenoid, bacterioruberin, extracted from Haloarcula japonica cells against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals was evaluated. The antioxidant capacity of bacterioruberin was much higher than that of β -carotene.
Haloarcula japonica, an extremely halophilic archaeon that requires high concentrations of NaCl for growth, accumulates the C 50 carotenoid bacterioruberin (BR). By homology analysis, a gene cluster, including c0507, c0506, and c0505, was found and predicted to be involved in the synthesis of bacterioruberin. To elucidate the function of the encoded enzymes, we constructed Ha. japonica mutants of these genes and analyzed carotenoids produced by the mutants. Our research showed that c0507, c0506, and c0505 encoded a carotenoid 3,4-desaturase (CrtD), a bifunctional lycopene elongase and 1,2-hydratase (LyeJ), and a C 50 carotenoid 2؆,3؆-hydratase (CruF), respectively. The above three carotenoid biosynthetic enzymes catalyze the reactions that convert lycopene to bacterioruberin in Ha. japonica. This is the first identification of functional CrtD and CruF in archaea and elucidation of the complete biosynthetic pathway of bacterioruberin from lycopene.
IMPORTANCEHaloarcula japonica, an extremely halophilic archaeon, accumulates the C 50 carotenoid bacterioruberin (BR). In this study, we have identified three BR biosynthetic enzymes and have elucidated their functions. Among them, two enzymes were found in an archaeon for the first time. Our results revealed the biosynthetic pathway responsible for production of BR in Ha. japonica and provide a basis for investigating carotenoid biosynthetic pathways in other extremely halophilic archaea. Elucidation of the carotenoid biosynthetic pathway in Ha. japonica may also prove useful for producing the C 50 carotenoid BR efficiently by employing genetically modified haloarchaeal strains.
Natural antimicrobial peptides (AMPs) are potential antibiotic alternatives. Marine crustaceans are thought to generate more powerful and various AMPs to protect themselves from infections caused by pathogenic microorganisms in their complex aquatic habitat, thus becoming one of the most promising sources of AMPs or other bioactive substances. In the study, a novel protein was identified as an interacting partner of male-specific AMP SCY2 in Scylla paramamosain and named scyreprocin. The recombinant product of scyreprocin (rScyreprocin) was successfully expressed in Escherichia coli. rScyreprocin exerted potent, broad-spectrum antifungal, antibacterial, and anti-biofilm activity (minimum inhibitory concentrations from 0.5 to 32 µM) through differential modes of action, including disruption of cell membrane integrity and induction of cell apoptosis, and has rapid bactericidal (in 0.5-2 h) and fungicidal (in 8-10 h) kinetics. In addition to its fungicidal activity against planktonic fungi, rScyreprocin also prevented the adhesion of fungal cells, inhibited biofilm formation, and eradicated the mature biofilms. Moreover, rScyreprocin showed a profound inhibitory effect on spore germination of Aspergillus spp. (minimum inhibitory concentrations from 4 to 8 µM). This peptide was not cytotoxic to murine and mammalian cells and could increase the survival rate of Oryzias melastigma under the challenge of Vibrio harveyi. Taken together, the novel AMP scyreprocin would be a promising alternative to antibiotics used in aquaculture and medicine.
Context:
Charcoal of Chinese drugs is a kind of special processing product in Chinese medicine and used for treatment of haemoptysis, hematemesis and haemorrhage in the clinic during ancient times. During carbonizing, significant changes occur in chemical constituents and the efficacy of haemostasis will be enhanced. But the quality control standard of ‘carbonizing retains characteristics’ should be followed.
Objective:
This review introduces the typical methods of carbonizing, which highlight current research progress on haemostatic substances of charcoal drugs so as to provide a reasonable explanation for the theory of haemostasis treated by charcoal medicine.
Methods:
English and Chinese literature from 2004 to 2019 was collected from databases including Web of Science, PubMed, Elsevier and CNKI (Chinese). Charcoal drug, chemical constituents, processing, haemostasis and carbon dots were used as the key words.
Results:
Charcoal drugs mainly play a haemostatic role and the effect can be classified into four types to stop bleeding: removing blood stasis, cooling blood, warming meridians and astringing. Changes in composition lead to changes in pharmacodynamics. Carbonizing methods and basic research on haemostasis material in charcoal drugs have also been summarized.
Conclusions:
This review summarizes the classification of charcoal drugs and highlights the possible material bases for the haemostatic effect of charcoal drugs in recent years, providing new insights to future research.
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