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A novel nanobody-drug conjugate (NDC) was constructed by incorporating an amphipathic peptide, GALA, which improved the cytotoxicity by one to two orders of magnitude. Mechanistic studies demonstrate that tethering to lipids induces GALA to form a helix, which dramatically enhances endocytosis. Our work provides a general strategy not only for improving the anti-cancer efficacy of protein-drug conjugates but also for increasing the efficiency of other types of endocytosis-dependent cell delivery.

Unique plant-derived cyclic peptides family exhibiting various key biological activities has great possibility for anticancer therapy. In this study, we investigated the effects of orbitides isolated from flax (

Emulsification is a key issue which makes oil hard to be separated during aqueous extraction from Camellia Oleifera Abel. seeds. Therefore, de-emulsification is required to recover and enhance oil yield. To investigate the major protein components resulting in a stable emulsion, the emulsifiable proteins of Camellia oleifera Abel. seeds (CSEP) were obtained from the emulsions formed during aqueous extraction of Camellia oil, and two major protein fractions (CSEP-A and CSEP-B) were isolated and purified from CSEP. CSEP-A consisted of a small amount of polysaccharides and seven major subunits/proteins with a molecular weight ranging from 13 to 35 kDa. CSEP-B was a protein mixture with a molecular weight range of 13–15 kDa. At neutral pH condition, CSEP-A had higher intrinsic fluorescence intensity and surface hydrophobicity compared to CSEP-B. There were more β-sheet, but lesser α-helix in protein secondary structure of CSEP-A. Both CSEP-A and CSEP-B were rich in Glu, Arg and Asp. However, hydrophobic amino acids contents in CSEP-A (33.47 ± 2.35%) was significantly higher (p

The aim of this study was to investigate the cellular uptake and antitumor activity of 2 different structural flaxseed orbitides ([1–9-NαC]-linusorb B2 containing methionine residue and [1–9-NαC]-linusorb B3 containing isoleucine residue) in SGC-7901 cells as well as its underlying molecule mechanism. Results showed that the absorption efficiency of [1–9-NαC]-linusorb B3 was higher than that of [1–9-NαC]-linusorb B2. Besides, through the evaluation of mitochondrial membrane potential (Δφm), cytochrome c (Cyt C) release, caspase, death receptor activation and cleavage of PARP, we found that [1–9-NαC]-linusorb B3 exhibited more obvious apoptosis induction on SGC-7901 cells than [1–9-NαC]-linusorb B2. Interestingly, [1–9-NαC]-linusorb B3-induced cell apoptosis might involve a mitochondrial- and death receptor-mediated intrinsic and extrinsic pathway; however, [1–9-NαC]-linusorb B2 induced cell apoptosis by the activation of Fas, FasL, caspase 8 as well as caspase 3 instead of Bax, Cyt C, tBid and caspase 9, suggesting that extrinsic pathway might be involved in the apoptotic process.

Mulberry leaves had been used as material of medicine, drink, and functional foods in many countries, but the variety suitability for different applications is still not clear. In this study, the nutritional and phytochemical components of mulberry tender shoots from 19 varieties in China were investigated. Obvious genotypic diversity was observed in all the assessed components. The contents of crude protein (CP), total soluble sugars (TSS), crude fiber (CF), 1-deoxynojirimycin (DNJ), total phenolic (TP), and total flavonoid content (TF) were 27.63–37.36 g/100 g dry weight (DW), 58.71–150.31 mg/g DW, 9.90–13.85 g/100 g DW, 0.08–1.12 mg/g DW, 8.76–20.26 mg gallic acid equivalents (GAE)/g DW, and 21.36–56.41 mg rutin equivalents (RE)/g DW, respectively. Chlorogenic acid, rutin, isoquercitrin, and astragalin were considered to be mainly potential antioxidant compounds in mulberry leaves. According to the result of correlational analysis, principal component analysis (PCA), and hierarchical cluster analysis (HCA), the NS14 variety had high comprehensive healthy properties. GS8, D10, XS, TP2, and DHS could be appropriate materials for functional food or drink because of high content of phenolics or DNJ. Some varieties may have a potential application as protein-rich vegetables. The study suggests that some variety of mulberry could be selected and utilized rationally for their dietary properties.

Medium- and long-chain triacylglycerols (MLCTs) with high 1,3-oleic-2-medium chain (OMO) triacylglycerols content were synthesized by the acidolysis reaction between Cinnamomum camphora seed oil (CCSO) and oleic acid, catalyzed by lipozyme RM IM. In addition, the simultaneous rate of acyl migration (RAM) was investigated during the esterification process. The maximal yield of OMO-structured triacylglycerols (YST) was up to 64.45% under the optimum condition: the molar ratio of CCSO to oleic acid, 1:4; enzyme dosage, 10 wt%; temperature, 60°C; and reaction time, 24 h. RAM was 28.66% at same time. It is shown that YST positively related to RAM at single experiments of the molar ratio (r2 = 0.999, P2 = 0.988, P2 = 0.923, P2 = 0.940, P2 = −0.682, P> .05) or a reaction time longer than 24 h (r2 = −0.594, P> .05). The effect of temperature on the reaction could be verified by molecular dynamics simulation, opened enzyme lids were increased with temperature elevated, and helped substrates invade into catalytic triad. When temperature rose to higher than 333 K (60°C), rearrangement of the catalytic triad was promoted, which resulted in both decreasing enzyme catalytic activity and irritating acyl migration. This study could help understand the trigger cause of acyl migration at high temperature, and conclusively reaction temperature was a vital factor to balance YST and RAM in a solvent-free system. OMO-structured triacylglycerols might possess a potential application as a new type of nutritional lipids for daily diet. Enzymatic Synthesis of 1,3-oleic-2-medium Chain Triacylglycerols and Strategy of Controlling Acyl Migration: Insights from Experiment and molecular dynamics simulation

A novel actinobacterium, designated strain JXJ CY 01T, was isolated from a mucilaginous sheath of Microcystis aeruginosa FACHB-905 collected from Lake Dianchi, south-west China. Taxonomic position of the isolate was determined by polyphasic approaches. Strain JXJ CY 01T shared 16S rRNA sequence similarities of 98.9 and 98.0% with Microbacterium marinilacus YM11-607T and Microbacterium paludicola US15T, and less than 98% with other members of the genus Microbacterium. The DNA–DNA relatedness values between strains JXJ CY 01T and M. marinilacus JCM 16546T and M. paludicola JCM 14308T were 53.5±1.4 and 53.8±2.1%, respectively. l-Ornithine was detected in the cell wall, and rhamnose, galactose, glucose, arabinose, fucose and mannose as signature sugars in the whole-cell hydrolysates. Other chemotaxonomic characteristics determined were MK-12 and MK-11 as predominant menaquinones, anteiso-C15:0, iso-C16:0, anteiso-C17:0 and iso-C17:0 as major cellular fatty acids (>10%), and diphosphatidylglycerol, phosphatidylglycerol, an unidentified glycolipid and two unidentified phospholipids as the polar lipids. The DNA G+C content was 70.4 mol%. On the basis of the above taxonomic data, strain JXJ CY 01T is determined to represent a novel species of the genus Microbacterium, for which the name Microbacterium lacusdiani sp. nov. is proposed. The type strain is JXJ CY 01T (= KCTC 29655T=DSM 29188T). The type strain JXJ CY 01T can solubilize both insoluble inorganic (calcium phosphate) and organic phosphate (l-α-phosphatidylcholine) and is possibly one of the mechanism for enhancement of growth of M. aeruginosa FACHB-905.

ArgR, a transcriptional regulator belonging to the AraC/XylS family, plays a key role in arginine metabolism regulation. ArgR has also been found to repress the transcription of a lipase gene, but its molecular mechanism is still unknown. In this study, we investigated the molecular mechanism acting on the expression of intracellular lipase gene lipA regulated by ArgR in Pseudomonas protegens Pf-5 through knockout and overexpression of argR, detection of DNA-protein interaction in vivo, determining whole-cell lipase activities of various strains derived from Pf-5, and examining β-galactosidase activities of various lacZ fusions. The results demonstrated that ArgR inhibits lipA expression at the transcriptional level. Further results showed that the inhibition of lipA transcription by ArgR is mediated by binding to the ArgR binding site of lipA promoter to produce steric hindrance, in which the common sequence, TGTCGC is crucial for the ArgR binding. Besides, arginine inhibits lipA expression in both wild-type and argR mutant, and shows a synergistic inhibition on lipA expression when combined with ArgR. To the best of our knowledge, this is the first report on ArgR directly repressing the transcription of a lipase gene.

Soybean dregs fermented by Neurospora crassa is a typical traditional food in Gannan district of China. In this study, in vitro imitated gut fermentation was carried out to evaluate whether the oligosaccharides from this fermented soybean dregs had potential prebiotic properties. 11.91% of oligosaccharides were extracted from the fermented soybean dregs at the optimized condition which of 1:25 for ratio of soybean dregs (g) to 50% ethanol (ml), 90 min of extracted duration at 70 °C for twice. The soybean dreg oligosaccharides (SBOS) were progressively purified with Sevag method and on columns filled with AB-8 macroporous resin, and then identified as cellobiose by HPLC-ESI-MS and FT-IR. Oligosaccharides of soybean dregs with 800 mg/L significantly decreased pH value (p

An antialgal compound was isolated from the cultured broth of Streptomyces jiujiangensis JXJ 0074(T) by using bioassay methods. Based on the data of (1)H-NMR, (13)C-NMR, ESI-MS, and thin layer chromatography, the active compound was identified as L-valine, which showed antialgal activity mainly against Microcystis. L-valine exhibited greater antialgal activities than both L-lysine and copper sulfate (CuSO4) did on Microcystis aeruginosa lawn. However, M. aeruginosa recovered growth earlier with higher growth rate in L-valine treatment than in L-lysine treatment. L-valine dissipated completely within 2 days, much quicker than L-lysine (6 days), which resulted in the lysing of more than 80 % M. aeruginosa cells and the release of amount of intracellular microcystin-LR (MC-LR) within 2 days. As a resultant, the extracellular MC-LR content was more than twice of the control from day 1 to 5. Exposure to L-valine significantly promoted the synthesis of MC-LR. L-lysine also promoted the release and synthesis of MC-LR with much lesser efficiency than L-valine. L-valine could damage Microcystis severely, causing perforation and collapse of M. aeruginosa cells and decrease of the chlorophyll. The superoxide dismutase (SOD) activity in L-valine-treated cells of M. aeruginosa initially increased with 32.94 ± 3.37 % higher than the control after 36 h and then decreased quickly. However, the increase rate of superoxide anion radical (O2 (-)) was much higher than that of SOD, which resulted in serious lipid peroxidation and accumulation of malondialdehyde (MDA). To our knowledge, this is the first report showing L-valine active against cyanobacteria.

Ginsenoside Rh2 (GRh2), isolated from Panax ginseng C. A. Meyer, has been proven as an anticancer compound both in vitro and in vivo. In the present study, we investigated the role of the lysosomes during the apoptosis of HepG2 cells induced by GRh2. The results showed that GRh2 significantly induced intracellular reactive oxygen species (ROS) generation in the HepG2 cells, which consequently resulted in early lysosomal membrane permeabilization with the release of cathepsin B (Cat B) to the cytosol. Western blot analysis showed that the released Cat B in the cytosol contributed to Bid cleavage. Subsequently mitochondrial damage was observed in the HepG2 cells. Interestingly, when the HepG2 cells were pre-treated with N-Acetyl-L-Cysteine (NAC) for 1 h, which inhibited ROS generation before being exposed to GRh2, the permeabilization of lysosomal membranes and the levels of Cat B in the cytosol were down-regulated. Moreover, mitochondrial damage was alleviated when the HepG2 cells were pre-treated with leupeptin (Leu). From the above results, it could be concluded that GRh2 induced apoptosis of the HepG2 cells through accumulation of ROS and activation of the lysosomal-mitochondrial apoptotic pathway involving the release of Cat B.

A novel actinobacterium, designated strain JXJ CY 21T, was isolated from the culture mass of Microcystis sp. FACHB-905 collected from Lake Dianchi, South-west China. Polyphasic taxonomic study revealed that the isolate should be a member of the genus Citricoccus. Comparison of the 16S rRNA gene sequence of strain JXJ CY 21T with the available sequences in the GenBank database showed that the strain is closely related to Citricoccus zhacaiensis FS24T (97.8 % similarity), Citricoccus parietis 02-Je-010T (97.7 %), Citricoccus terreus V3M1T (97.6 %), Citricoccus nitrophenolicus PNP1T (97.2 %), Citricoccus alkalitolerans YIM 70010T (97.2 %) and Citricoccus muralis 4-0T (97.0 %). The DNA–DNA hybridization values between strain JXJ CY 21T and the related type strains C. zhacaiensis FS24T and C. parietis 02-Je-010T were 16.0 ± 2.6 and 5.4 ± 1.7 %, respectively. The peptidoglycan in the cell wall was A4α type containing lysine–glutamic acid–glycine. The major respiratory menaquinone was found to be MK-8 (H2) (98.5 %), while the major cellular fatty acids (>10 %) were anteiso-C15:0, iso-C16:0, iso-C15:0 and iso-C14:0. The polar lipids detected were diphosphatidylglycerol, phosphatidylinositol, phosphatidylglycerol, an unidentified phospholipid and an unidentified glycolipid. The DNA G + C content was determined to be 62.7 mol%. Strain JXJ CY 21T can solubilize both insoluble inorganic and organic phosphates up to 24.7 and 1.7 mg/l respectively. This property of the novel actinobacterium acts as a modulator for enhancement of growth of Microcystis sp. FACHB-905 in the lake ecosystem where the amount of soluble phosphate is limited. On the basis of the above taxonomic data, strain JXJ CY 21T represents a novel species of the genus Citricoccus, for which the name Citricoccus lacusdiani sp. nov. is proposed. The type strain is JXJ CY 21T (=KCTC 29653T = DSM 29160T).

Copper sulfate (CuSO 4 ) has been widely used as an algicide to control harmful cyanobacterial blooms (CyanoHABs) in freshwater lakes. However, there are increasing concerns about this application, due mainly to the general toxicity of CuSO 4 to other aquatic species and its long-term persistence in the environment. This study reported the isolation and characterization of two natural algicidal compounds, i.e., tryptamine and tryptoline, from Streptomyces eurocidicus JXJ-0089. At a concentration of 5 μg/ml, both compounds showed higher algicidal efficiencies than CuSO 4 on Microcystis sp. FACHB-905 and some other harmful cyanobacterial strains. Tryptamine and tryptoline treatments induced a degradation of chlorophyll and cell walls of cyanobacteria. These two compounds also significantly increased the intracellular oxidant content, i.e., superoxide anion radical (O 2 − ) and malondialdehyde (MDA), but reduced the activity of intracellular reductants, i.e., superoxide dismutase (SOD), of cyanobacteria. Moreover, tryptamine and tryptoline treatments significantly altered the internal and external contents of microcystin-LR (MC-LR), a common cyanotoxin. Like CuSO 4 , tryptamine and tryptoline led to releases of intracellular MC-LR from Microcystis , but with lower rates than CuSO 4 . Tryptamine and tryptoline (5 μg/ml) in cyanobacterial cultures were completely degraded within 8 days, while CuSO 4 persisted for months. Overall, our results suggest that tryptamine and tryptoline could potentially serve as more efficient and environmentally friendly alternative algicides than CuSO 4 in controlling harmful cyanobacterial blooms. IMPORTANCE Cyanobacterial harmful algal blooms (CyanoHABs) in aquatic environments have become a worldwide problem. Numerous efforts have been made to seek means to prevent, control, and mitigate CyanoHABs. Copper sulfate (CuSO 4 ), was once a common algicide to treat and control CyanoHABs. However, its application has become limited due to concerns about its general toxicity to other aquatic species and its long-term persistence in the environment. There is a great need for algicides with higher specificity and low environmental impacts. This study reports the isolation and characterization of two natural algicidal compounds from a streptomycete strain, Streptomyces eurocidicus JXJ-0089. Our results suggest that the identified algicides could potentially serve as more efficient and environmentally friendly alternative algicides than CuSO 4 in controlling harmful cyanobacterial blooms.

A novel actinobacterium, designated strain JXJ CY 19T, was isolated from a culture mat of Microcystis aeruginosa FACHB-905 collected from Dianchi Lake, South-west China. 16S rRNA gene sequences comparison of strain JXJ CY 19T and the available sequences in the GenBank database showed that the strain was closely related to Modestobacter marinus 42H12-1T (99.1% similarity) and Modestobacter roseus KLBMP 1279T (99.0%). The isolate had meso-diaminopimelic in the cell wall with whole-cell sugars of mannose, rhamnose, ribose, glucose, galactose, and arabinose. The menaquinone detected was MK-9(H4), while the major cellular fatty acids include C17:1 ω8c, C15:0 iso, C15:1 iso G and C16:0 iso. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and an unidentified phospholipid. The DNA-DNA hybridization values between strains JXJ CY 19T and the closely related type strains Modestobacter marinus CGMCC 4.5581T and Modestobacter roseus NBRC 108673T were determined to be 50.8 ± 0.8% and 44.1 ± 1.7%, respectively. The DNA G+C content was 71.9 mol%. On the basis of the above taxonomic data and differences in physiological characters from the closely related type strains, strain JXJ CY 19T was recognized as a novel species of the genus Modestobacter, for which the name Modestobacter lacusdianchii sp. nov. (JXJ CY 19T = KCTC 39600T = CPCC 204352T) is proposed. The type strain JXJ CY 19T can solubilize calcium phosphate tribasic (Ca3(PO4)2), phytin and L-α-phosphatidylcholine. The phosphate-solubilizing property of the novel actinobacterium could be a possible factor for the increase in growth of Microcystis aeruginosa FACHB-905 in ecosystem where the amount of available soluble phosphate is limited such as Dianchi Lake.

Strain JXJ 0135(T), an anti-cyanobacterial actinomycete, was isolated from a soil sample collected from Lushan Mountain, south China, and identified by using polyphasic approach. Phylogenetic analysis of the near-complete 16S rRNA gene sequence indicated that strain JXJ 0135(T) belongs to the genus Streptomyces and exhibited distinct subclade and also highest similarity (98.6%) to Streptomyces scopuliridis RB72(T). The strain developed well-branched substrate and aerial mycelia, and produced spiral spore chains. Spores were elliptical and the spore surface was smooth. The strain contained LL-diaminopimelic acid with whole-cell sugars of mannose, rhamnose, glucose and galactose. Phospholipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannosides, phosphotidylinositol dimannoside, an unidentified aminophospholipid and an unknown phospholipid. The menaquinones were MK-9(H6) and MK-9(H8). The major components of the fatty acids were anteiso-C15:0, iso-C16:0, anteiso-C17:0, iso-C15:0, C16:0, iso-C17:0 and iso-C14:0. The G+C content was 69.3 mol%. The DNA-DNA hybridization value between JXJ 0135(T) and S. scopuliridis RB72(T) was 41.2±1.4%. On the basis of the polyphasic data, strain JXJ 0135(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces lushanensis sp. nov. is proposed. The type strain is JXJ 0135(T) (=DSM 42121(T)=JCM 19628(T)=KCTC 29261(T)=KACC 17834(T)=NRRL B-24994(T)).

An actinomycete capable of lysing cyanobacteria, strain JXJ 0074(T), was isolated from a soil sample collected from Jiangxi province, south China, and characterized by using polyphasic taxonomy. The new isolate showed morphological and chemotaxonomic properties typical of members of the genus Streptomyces. Phylogenetic analysis of the near-complete 16S rRNA gene sequence indicated that strain JXJ 0074(T) should be affiliated to the genus Streptomyces and exhibited highest similarities to Streptomyces shenzhenensis DSM 42034(T) (98.99 %) and Streptomyces lucensis NBRC 13056(T) (98.60 %), while the similarities to other members of the genus are lower than 98.22 % similarity. However, the DNA-DNA hybridization values between strain JXJ 0074(T) and S. shenzhenensis DSM 42034(T) or S. lucensis NBRC 13056(T) were 46.2 ± 2.6 and 32.6 ± 3.1 %, respectively. Thus, on the basis of the polyphasic data, strain JXJ 0074(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces jiujiangensis sp. nov. is proposed. The type strain is JXJ 0074(T) (= BCRC 16953(T) = KCTC 29262(T)).