ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (11), pp. 001-009, November, 2018. © International Scholars Journals
Full Length Research Paper
Expression and identification of a small recombinant beefy meaty peptide secreted by the methylotrophic yeast Pichia pastoris
Song-Rong Zeng1,2, Yan-Ping Wang1*, Yong-Qing Yang1,3 and Jian Zhang1
1College of Food Engineering and Biotechnology, Key Laboratory of Food Nutrition and Safety of the Ministry of Education, Tianjin Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, P. R. China.
2Yingdong College of Life Sciences, Shaoguan University, Shaoguan 512005, P. R. China.
3Handan Municipal Center for Disease Prevention and Control, Handan 056002, P. R. China.
Accepted 10 November, 2018
Abstract
A tasty peptide, beefy meaty peptide (BMP), which was initially isolated from beef digested by papain, has potential as a new flavor enhancer. BMP is a small peptide consisting of eight amino acids. In this study, DNA encoding BMP was synthesized and cloned into the expression vector pPICZ A to obtain the recombinant expression vector pPICZ A-BMP. The recombinant vector was linearized and then integrated into the genome of Pichia pastoris strain X-33 by electroporation. The recombinant strain P.p2 was then incubated in BMGY medium and then induced in BSM medium for expression of BMP. The fermentation broth was centrifuged and the supernatant was purified by gel-filtration chromatography. Purified recombinant BMP was then tested by high performance liquid chromatography- mass spectrometry (HPLC-MS). After optimization of the culturing process, the yield of BMP reached 10 mg/L in the clarified broth. The results of mass spectrometry/mass spectrometry (MS/MS) showed that recombinant BMP secreted from P. pastoris had a molecular weight of 873.7 Da rather than the expected 847 Da. We speculate that the last amino acid, Ala, of the BMP sequence was replaced with Pro in the BMP fragment.
Key words: beefy meaty peptide, Pichia pastoris, peptide expression, HPLC-MS.
Yan-Ping Wang*, Song-Rong Zeng, Yong-Qing Yang and Jian Zhang
Page: 1 - 9
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (11), pp. 001-011, November, 2018. © International Scholars Journals
Full Length Research Paper
Comparative analyses of phytochemicals and antimicrobial properties of extracts of wild Tamarindus indica pulps
Adeola, A.A1*, Adeola, O.O.2 and Dosumu, O.O.3
1Department of Home Economics, Emmanuel Alayande College of Education, Oyo, Nigeria.
2Department of Chemistry, University of Ibadan, Ibadan, Nigeria.
3Department of Chemistry, University of Ilorin, Ilorin, Nigeria.
Accepted 4 November, 2018
Abstract
Tamarindus indica L. (Tamarind), an underutilised fruit tree which belongs to the Leguminosae family, grows wild in the savannah region of Nigeria. T. indica pulp was obtained from 19 towns of the 20 savannah states of Nigeria. The methanol and hexane crude extracts obtained from it pulps were evaluated in vitro to determine their inhibition activities on human pathogenic microorganisms made up of five bacteria and three fungi. All the bacterial strains were sensitive to both extracts at concentrations ranging from 25 to 125 mg/ml, using the agar broth cup diffusion procedure. Only the hexane extract exhibited intrinsic antifungal properties on Penicillium species. Preliminary phytochemical screening of both extracts indicated the presence of alkaloids and tannins. Both the antimicrobial and phytochemical properties of the extracts of the pulp varied for locations of the tamarind. Natural products present in tamarind pulp have potential of being used as agents for animals and/or plants protector against pathogenic microorganisms.
Key words: Tamarind, phytochemical, antimicrobial, underutilised fruit, saponins.
A.A* , O.O and Dosumu, Adeola , Adeola , O.O
Page: 1 - 11
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (11), pp. 001-003, November, 2018. © International Scholars Journals
Full Length Research Paper
Nested PCR on semen samples for the detection of Mycobacterium avium subsp paratuberculosis
Ali Sharifzadeh1*, Abbas Doosti2, Mohammad Hashem Fazeli1 and Iman Adavoudi3
1Department of Microbiology, Faculty of Veterinary Medicine, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran.
2Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran.
3Faculty of Veterinary Medicine, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran.
Accepted 8 November, 2018
Abstract
Mycobacterium paratuberculosis (MAP) is a gram positive, acid-fast bacterium and cause of Johne’s disease in some animals. The important signs of this disease in bovine are diarrhea, weight loss, bowel inflammation, fever and reduce of milk production. The symptoms of this disease are very similar to Crohn©s disease in humans. The aim of this study was to use nested -PCR as an accurate and fast method to trace MAP in bull semen. Semen samples from 112 bulls were collected and DNA was extracted. Then, nested-PCR was performed by specific primers for IS900 gene of MAP. The PCR products with 230 bp length were estimated as a positive. The frequency of MAP in semen samples were 12.50%. The results were showed nested-PCR is a good procedure with high efficiency for detection of intracellular bacteria such as M. paratuberculosis in bull©s semen samples. Thus, despite this abundance more attention to this disease in bulls to identify MAP quickly is essential.
Key words: MAP, nested-PCR, John’s disease, IS900 gene, semen, bull.
Mohammad Hashem Fazeli and Iman Adavoudi, Abbas Doosti, Ali Sharifzadeh*
Page: 1 - 3
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (11), pp. 001-005, November, 2018. © International Scholars Journals
Full Length Research Paper
Prevalence of plasmid-mediated 16S rRNA methylase genes among Proteus mirabilis isolates from a Chinese hospital
Fangyou Yu1, Jingye Pan2, Baixing Ding2, Lehe Yang3, Xueqing Zhang1 and Liangxing Wang3*
1Department of Laboratory Medicine, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou.325000, China.
2Department of Intensive Care Unit, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou.325000, China.
3Department of Respiratory Medicine, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou.325000, China.
Accepted 11 November, 2018
Abstract
16S rRNA methylase-mediated high-level resistance to aminoglycosides has been reported recently in clinical isolates of Gram -negative bacilli from several countries. Five (2.5%, 5/198) of 198 isolates of Proteus mirabilis from a teaching hospital in Wenzhou, China, were positive for 16S rRNA methylase genes (one for armA, four for rmtB) and highly resistant to gentamicin, amikacin and tobramycin (MICs, 256 g/ml). One of five isolates harboring 16S rRNA methyalse genes were extended-spectrum - lactamases (ESBL) producer. The plasmids harboring 16S rRNA methylase genes from four out of five donors were transferred into the recipients, Escherichia coli J53. Among five isolates harboring armA and rmtB, the armA gene and the rmtB genes were located on the plasmids, as determined by Southern hybridization. The present study investigated the prevalence of 16S rRNA methylase genes in clinical isolates of P. mirabilis in China for the first time.
Key words: Proteus mirabilis, 16S rRNA methylase, plasmid.
Fangyou Yu, Baixing Ding, Jingye Pan, Xueqing Zhang and Liangxing Wang*, Lehe Yang
Page: 1 - 5
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (11), pp. 001-009, November, 2018. © International Scholars Journals
Full Length Research Paper
Nanomolecular simulation of the voltage–gated potassium channel protein by gyration radius study
F. Mollaamin1*, I. Layali2, A. R. Ilkhani3 and M. Monajjemi1
1Department of Chemistry, Science and Research Branch, Islamic Azad University, Tehran, Iran.
2Science and Research Branch, Islamic Azad University, Tehran, Iran.
3Department of Chemistry, Yazd Branch, Islamic Azad University, Yazd, Iran.
Accepted 23 July, 2018
Abstract
Molecular dynamics simulations may be used to probe the interactions of membrane proteins with lipids and with detergents at atomic resolution. Examples of such simulations for ion channels and for bacterial outer membrane have already been studied. The molecular potassium channel function is universally conserved. Potassium channels allow potassium flux and are essential for the generation of electric current across excitable membranes. Potassium channels are also the targets of various intracellular control mechanisms, such that the suboptimal regulation of channel function might be related to pathological conditions. Realistic studies of ion current in biologic channels, present a major challenge for computer simulation approaches. In this work, to characterize protein behavior, we observed quantities such as gyration radius and energy average. We studied the changes of these factors for voltage – gated potassium channel protein in gas phase with native conformation by Monte Carlo, Molecular and Langevin Dynamics simulations. Monte Carlo simulation is a stochastic method and therefore, is the best method to evaluate the radius of gyration. When the temperature is increased the kinetic energy is increased too and its correlation is linear. All the calculations were carried out By Hyperchem 8.0 program. The determination of gyration radius is spectacular for configuration of a macromolecule. It also reflects molecular compactness shape. The radius of gyration is calculated by VMD 1.8.7 software.
Key words: Monte Carlo simulation, molecular dynamics simulation, Langevin dynamics simulation, protein folding, gyration radius.
I. Layali, A. R. Ilkhani and M. Monajjemi, F. Mollaamin*
Page: 1 - 9
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (11), pp. 001-007, November, 2018. © International Scholars Journals
Full Length Research Paper
Rapid detection of community acquired-methicillin resistance Staphylococcus aureus recovered from King Saudi Arabia
Moussa I. M.1* and Hessan A. M.2
1Center of Excellence in Biotechnology Research, King Saud University, P. O. Box 2460, Riyadh 11451, Saudi Arabia.
2College of Applied Studies and Community Service, King Saud University, P. O. Box 2460 Riyadh, King Saudi Arabia.
Accepted 06 July, 2018
Abstract
The objectives of this study were to examine methicillin-resistant Staphylococcus aureus (MRSA) strains recovered from major hospitals in King Saudi Arabia (KSA) to determine the percent of community acquired MRSA (CA-MRSA) phenotypically by conventional methods and genotypically by multiplex polymerase chain reaction (multiplex-PCR) for direct and simultaneous detection of S. aureus 16S rRNA, Panton–valentine leucocidin (PVL ) and staphylococcal cassette chromosome mec (SCCmec) type IVa genes. Therefore, 135 strains of S. aureus collected during the period of 2008 and 2009 from major hospital laboratories and public health centers, Riyadh, King Saudi Arabia were tested phenotypically by conventional methods and genotypically by multiplex-PCR. PCR enables rapid detection of all 135 bacterioloically identified S. aureus (100%) as well as the mecA gene in all strains phenotypically resistant to methicillin (100%) . Moreover, it could detect the mecA gene in 8 strains (6%) phenotypically sensitive to methicillin. Only 18 strains (13.33%) recovered from skin and soft tissue infections were positive for PVL and (SCCmec) type IV. The results of this study indicate that the incidence of patients with CA-MRSA disease had been increased in Riyadh, KSA.
Key words: Staphylococcus aureus, CA-MRSA, PVL gene, multiplex polymerase chain reaction.
Hessan A. M., Moussa I. M*
Page: 1 - 7