Production of Human Papillomavirus 6b L1 Virus-like Particles Incorporated with Enhanced Green Fluorescent Whole Protein in Silkworm Larvae

Palaniyandi Muthukutty,Tatsuya Kato,Enoch Y. Park 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.3

Using human papillomavirus (HPV) as a subunit vaccine and its manipulation of surface loops is current trending research. Since the atomic model of L1 protein conformations were deciphered, their manipulations of epitopes bring multivalent vaccines. Here, in the present study, we have manipulated antigenic loops of HPV 6b L1capsid proteins in the amino acid regions 174 ~ 175(L1:174EGFP) and 348 ~ 349 (L1:348EGFP) with whole enhanced green fluorescent protein(EGFP), expressed in the silkworm larva using Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid technology. The expressed proteins were partially purified using sucrose density-gradient centrifugation and size-exclusion chromatography (SEC). The display of EGFP in virus-like particles (VLPs) was confirmed by immuno-fluorescence microscopy, Western blots and immune-transmission electron microscopy (immuno-TEM). There was higher expression of EGFP incorporated L1:174EGFP than L1:348EGFP. Hydrodynamic diameter of VLPs was corroborated by dynamic light scattering,confirming the size of expected range of around 160 nm and substantiating the incorporation of EGFP. From immuno-TEM, each L1:EGFP VLP formed small particles,suggesting that small particles of L1:EGFP fusion protein were aggregated. Our study illustrates that incorporation of whole protein can efficiently form chimeric VLPs, without hindering the conformation. HPV L1 protein accommodated a whole protein on its antigenic loop as a small particle, but an inserted whole protein was unstable.

Preparation of Ginseng Extract with Enhanced Levels of Ginsenosides Rg1 and Rb1 Using High Hydrostatic Pressure and Polysaccharide Hydrolases

Palaniyandi, Sasikumar Arunachalam,Suh, Joo-Won,Yang, Seung Hwan Medknow PublicationsMedia Pvt Ltd 2017 Pharmacognosy magazine Vol.13 No.49

<P><B>Background:</B></P><P>Ginsenosides are the principal components responsible for the pharmacological activities of ginseng. Ginsenosides Rg1 and Rb1 are the major compounds recognized as marker substances for quality control of ginseng-based products. These major compounds can be transformed to several pharmacologically active minor ginsenosides by chemical, microbial, and enzymatic means.</P><P><B>Materials and Methods:</B></P><P>In the present study, a combination of polysaccharide hydrolases and high hydrostatic pressure (HHP) were used to extract ginseng saponins enriched with ginsenosides Rg1 and Rb1. Temperature, pH, time, ginseng-to-water ratio, and pressure were optimized to obtain the maximum amount of Rg1 and Rb1 in the resulting extract using commercial polysaccharide hydrolases.</P><P><B>Results:</B></P><P>This study showed that treatment with a combination of cellulase, amylase, and pectinase at 100 MPa pressure, pH 4.8, and 45°C for 12 h resulted in higher Rg1 and Rb1 levels in the extract.</P><P><B>Conclusion:</B></P><P>This study describes a cheap and ecofriendly method for preparing ginseng extract enriched with Rg1 and Rb1.</P><P><B>SUMMARY</B></P><P><P>Ginsenosides are the principal bioactive components present in ginseng</P><P>Ginsenosides Rg1 and Rb1 are the most abundant compounds in ginseng</P><P>High hydrostatic pressure (HHP) and Polysaccharide hydrolases (PH) were combined to extract ginseng saponins enriched with Rg1 and Rb1</P><P>Extraction conditions were optimized to obtain the maximum amount of Rg1 and Rb1</P><P>Extraction with a combination of cellulase, amylase, and pectinase at 100 MPa pressure at pH 4.8, and 45°C for 12 h resulted in higher levels of Rg1 and Rb1 in the ginseng extract</P></P> >[FIG OMISSION]</BR><P><B>Abbreviations used:</B> ATCC: American Type Culture Collection, Mpa: Mega Pascal</P>

Effects of actinobacteria on plant disease suppression and growth promotion

Palaniyandi, Sasikumar Arunachalam,Yang, Seung Hwan,Zhang, Lixin,Suh, Joo-Won Springer-Verlag 2013 Applied microbiology and biotechnology Vol.97 No.22

<P>Biological control and plant growth promotion by plant beneficial microbes has been viewed as an alternative to the use of chemical pesticides and fertilizers. Bacteria and fungi that are naturally associated with plants and have a beneficial effect on plant growth by the alleviation of biotic and abiotic stresses were isolated and developed into biocontrol (BCA) and plant growth-promoting agents (PGPA). Actinobacteria are a group of important plant-associated spore-forming bacteria, which have been studied for their biocontrol, plant growth promotion, and interaction with plants. This review summarizes the effects of actinobacteria as BCA, PGPA, and its beneficial associations with plants.</P>

Biological control of anthracnose (<i>Colletotrichum gloeosporioides</i>) in yam by <i>Streptomyces</i> sp.MJM5763

Palaniyandi, S.A.,Yang, S.H.,Cheng, J.H.,Meng, L.,Suh, J.‐,W. Blackwell Publishing Ltd 2011 Journal of applied microbiology Vol.111 No.2

<P><B>Abstract</B></P><P><B>Aim: </B> To find a suitable biocontrol agent for yam anthracnose caused by <I>Colletotrichum gloeosporioides</I>.</P><P><B>Methods and Results: </B> An actinobacterial strain, MJM5763, showing strong antifungal activity, multiple biocontrol and plant growth‐promoting traits was isolated from a yam cultivation field in Yeoju, South Korea. Based on morphological and physiological characteristics and analysis of the 16S rDNA sequence, strain MJM5763 was identified as a novel strain of <I>Streptomyces</I> and was designated as <I>Streptomyces</I> sp. MJM5763. Treatment with MJM5763 and the crude culture filtrate extract (CCFE) was effective in suppressing anthracnose in detached yam leaves <I>in vitro</I> and reduced incidence and severity of anthracnose in yam plants under greenhouse conditions. The CCFE treatment was the most effective of all the treatments and reduced the anthracnose severity by 85–88% and the incidence by 79–81%, 90 days after inoculation with the pathogen. CCFE treatment was also effective under field conditions and showed a reduction of 86 and 75% of anthracnose severity and incidence, respectively.</P><P><B>Conclusion: </B> <I>Streptomyces</I> sp. strain MJM5763 was effective in biocontrolling anthracnose in yam caused by <I>C.?gloeosporioides</I>.</P><P><B>Significance and Impact of the Study: </B> <I>Streptomyces</I> sp. MJM5763 is a potential alternative to chemical fungicides for reducing yield losses to anthracnose in yam.</P>

Bio-Inspired Green Nanoparticles: Synthesis, Mechanism, and Antibacterial Application

Palaniyandi Velusamy,Govindarajan Venkat Kumar,Venkadapathi Jeyanthi,Jayabrata Das,Raman Pachaiappan 한국독성학회 2016 Toxicological Research Vol.32 No.2

In the recent years, noble nanoparticles have attracted and emerged in the field of biology, medicine and electronics due to their incredible applications. There were several methods have been used for synthesis of nanoparticles such as toxic chemicals and high energy physical procedures. To overcome these, biological method has been used for the synthesis of various metal nanoparticles. Among the nanoparticles, silver nanoparticles (AgNPs) have received much attention in various fields, such as antimicrobial activity, therapeutics, bio-molecular detection, silver nanocoated medical devices and optical receptor. Moreover, the biological approach, in particular the usage of natural organisms has offered a reliable, simple, nontoxic and environmental friendly method. Hence, the current article is focused on the biological synthesis of silver nanoparticles and their application in the biomedical field.

Molecular cloning and characterization of the ABA-specific glucosyltransferase gene from bean (Phaseolus vulgaris L.)

Palaniyandi, S.A.,Chung, G.,Kim, S.H.,Yang, S.H. G. Fischer 2015 Journal of plant physiology Vol.178 No.-

Levels of the plant hormone abscisic acid (ABA) are maintained in homeostasis by a balance of its biosynthesis, catabolism and conjugation. The detailed molecular and signaling events leading to strict homeostasis are not completely understood in crop plants. In this study, we obtained cDNA of an ABA-inducible, ABA-specific UDP-glucosyltransferase (ABAGT) from the bean plant (Phaseolus vulgaris L.) involved in conjugation of a glucose residue to ABA to form inactive ABA-glucose ester (ABA-GE) to examine its role during development and abiotic stress in bean. The bacterially expressed PvABAGTase enzyme showed ABA-specific glucosylation activity in vitro. A higher level of the PvABAGT transcript was observed in mature leaves, mature flowers, roots, seed coats and embryos as well as upon rehydration following a period of dehydration. Overexpression of 35S::PvABAGT in Arabidopsis showed reduced sensitivity to ABA compared with WT. The transgenic plants showed a high level of ABA-GE without significant decrease in the level of ABA compared with the wild type (WT) during dehydration stress. Upon rehydration, the levels of ABA and phaseic acid (PA) decreased in the WT and the PvABAGT-overexpressing lines with high levels of ABA-GE only in the transgenic plants. Our findings suggest that the PvABAGT gene could play a role in ABA homeostasis during development and stress responses in bean and its overexpression in Arabidopsis did not alter ABA homeostasis during dehydration stress.

In Vitro Characterization of Lactobacillus plantarum Strains with Inhibitory Activity on Enteropathogens for Use as Potential Animal Probiotics

Palaniyandi, S. A.,Damodharan, K.,Suh, J. W.,Yang, S. H. Association of Microbiologists of India 2017 Indian journal of microbiology Vol.57 No.2

<P>The present study evaluates the probiotic properties of three Lactobacillus plantarum strains MJM60319, MJM60298, and MJM60399 possessing antimicrobial activity against animal enteric pathogens. The three strains did not show bioamine production, mucinolytic and hemolytic activity and were susceptible to common antibiotics. The L. plantarum strains survived well in the simulated orogastrointestinal transit condition and showed adherence to Caco-2 cells in vitro. The L. plantarum strains showed strong antimicrobial activity against enterotoxigenic Escherichia coli, Shiga toxin-producing E. coli, Salmonella enterica subsp. enterica serovar Typhimurium, Choleraesuis and Gallinarum compared to the commercial probiotic strain Lactobacillus rhamnosus GG. The mechanism of antimicrobial activity of the L. plantarum strains appeared to be by the production of lactic acid. Furthermore, the L. plantarum strains tolerated freeze-drying and maintained higher viability in the presence of cryoprotectants than without cryoprotectants. Finally, the three L. plantarum strains tolerated NaCl up to 8% and maintained > 60% growth. These characteristics of the three L. plantarum strains indicate that they could be applied as animal probiotic after appropriate in vivo studies.</P>

Bio-Inspired Green Nanoparticles: Synthesis, Mechanism, and Antibacterial Application

Velusamy, Palaniyandi,Kumar, Govindarajan Venkat,Jeyanthi, Venkadapathi,Das, Jayabrata,Pachaiappan, Raman Korean Society of ToxicologyKorea Environmental Mu 2016 Toxicological Research Vol.32 No.2

In the recent years, noble nanoparticles have attracted and emerged in the field of biology, medicine and electronics due to their incredible applications. There were several methods have been used for synthesis of nanoparticles such as toxic chemicals and high energy physical procedures. To overcome these, biological method has been used for the synthesis of various metal nanoparticles. Among the nanoparticles, silver nanoparticles (AgNPs) have received much attention in various fields, such as antimicrobial activity, therapeutics, bio-molecular detection, silver nanocoated medical devices and optical receptor. Moreover, the biological approach, in particular the usage of natural organisms has offered a reliable, simple, nontoxic and environmental friendly method. Hence, the current article is focused on the biological synthesis of silver nanoparticles and their application in the biomedical field.

Fermentative transformation of ginsenosides by a combination of probiotic Lactobacillus helveticus and Pediococcus pentosaceus

Sasikumar Arunachalam Palaniyandi,Bao Le,김진만,양승환 한국미생물학회 2018 미생물학회지 Vol.54 No.4

Ginseng are native traditional herbs, which exhibit excellentpharmacological activities. Probiotic Lactobacillus helveticusKII13 and Pediococcus pentosaceus strain KID7 were used forginsenoside transformation by fermenting crude ginseng extractto enhance minor gisenoside content. Thin-layer chromatography(TLC) analysis of fermented ginseng extract showed that theminor ginsenosides Rg3, Rh1, and Rh2 were main productsafter 5 days of fermentation. HPLC analysis was performed toquantify the major and minor ginsenosides. The Rg3 peakappeared on the 3rd day while the appearance of Rh2 peak andRh1 peak were observed on the 5th day. The co-culture of L. helveticus KII13 and P. pentosaceus KID7 converted majorginsenosides (Rb1 and Rg1) into minor ginsenosides (Rg3,Rh2, and Rh1).

Fermentative Transformation of Ginsenoside Rb1 from Panax ginseng C. A. Meyer to Rg3 and Rh2 by Lactobacillus paracasei subsp. tolerans MJM60396

Sasikumar Arunachalam Palaniyandi,손병모,Karthiyaini Damodharan,서주원,양승환 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.5

Lactic acid bacteria (LAB) were screened for ginsenoside transforming activity using crude ginseng extract. Thin-layer chromatography analysis of fermented ginseng extract showed that LAB strain MJM60396 possessed higher ginsenoside transformation ability than other strains. It converted major ginsenosides into minor ginsenosides such as Rg3 and Rh2. MJM60396 also showed high β-glucosidase activity. Strain MJM60396 was identified as Lactobacillus paracasei subsp. tolerans based on 16S rRNA gene sequence. To delineate the pathway involved in the production of the minor ginsenosides Rg3 and Rh2, strain MJM60396 was incubated with pure ginsenoside Rb1. HPLC analysis revealed the appearance of Rg3 and Rh2 peak from the incubation mixture containing Rb1 and strain MJM60396. Furthermore, β-glucosidase enzyme was prepared from strain MJM60396. To achieve its maximum activity, we optimized the pH and temperature conditions. Cell-free β-glucosidase enzyme hydrolyzed ginsenoside Rb1 through the following pathway: ginsenoside Rb1 → Rd → Rg3 → Rh2. This is the first report on the transformation of ginsenosides Rb1 to Rg3 and Rh2 by a Lac. paracasei subsp. tolerans strain. Our results indicate that Lac. paracasei subsp. tolerans MJM60396 has the potential to be used for preparing ginsenosides Rg3 and Rh2 as nutraceuticals.