• Journal of Microbiology and Biotechnology
• /
• v.24 no.5
• /
• pp.619-628
• /
• 2014

To identify lipase LipA (PFL_0617) from Pseudomonas protegens Pf-5, a lipA deletion mutant (Pf0617) and a complementary strain (Pf0617lipA) were constructed, and their effects on the lipase production were examined. Pf0617 remarkably decreased its whole-cell lipase activity, whereas Pf0617lipA made its whole-cell lipase activity not only restore to wild-type level but also get a further increment. However, the deletion and overexpression of lipA did not affect the extracellular lipase activity. In addition, the unbroken whole cells of these strains were able to catalyze the hydrolysis of membrane-permeable p-nitrophenyl esters, but could not hydrolyze the membrane-impermeable olive oil. These results confirmed that LipA was an intracellular lipase and Pf-5 could also be used as a natural whole-cell biocatalyst. To evaluate the potential of Pf-5 as a whole-cell biocatalyst and separately characterize the whole-cell LipA, the properties of the whole-cell lipases from Pf-5 and Top10lipA were characterized. The results demonstrated that both Pf-5 and Top10lipA exhibited high tolerance to alkaline condition, high temperature, heavy metal ions, surfactants, and organic solvents. Taken together, lipA can realize functional expression in E. coli Top10, and Pf-5 and Top10lipA as whole-cell biocatalysts may have enormous potential in applications.

• KSBB Journal
• /
• v.14 no.3
• /
• pp.264-272
• /
• 1999

Benzoylformate was converted to benzaldehyde by whole cell enzyme from Pseudomonas putida KCTC 1751. We investigated the effect of the composition of the growth medium on th accumulation of benzoylformate decarboxylase in the microbial cell. We prepared a calcium alginate capsule containing Pseudomonas putida cells to develop a reusable whole cell enzyme. Pseudomonas putida cells were inoculated in the capsule and cultured in M1 medium for 1 day followed by cultivation in M3 medium for 3 days. The dry cell density reached 77.75 g/L on the basis of the inner volume of the capsule. The specific activity of encapsulated whole cell benzoylformate decarboxylase was half as high as that of free whole cell enzyme. The activity of encapsulated whole cell benzoylformate decarboxylase was half as high as that of free whole cell enzyme. The activity of encapsulated whole cell benzoylformate decarboxylase decreased 20 % after use for 20 batches and 40% after use for 30 batches. The dry cell density reduced about 10 % after 30 trials.

• Journal of Plant Biology
• /
• v.19 no.3
• /
• pp.71-84
• /
• 1976

In order to observe the phosphate metabolism in chloroplast, the contents of inorganic phosphate and various compounds in chloroplast from spinach leaf tissues were investigated during the reaction in the light and dark in the reaction mixture and the turnover of phosphate in chloroplast was compared with that of whole cell system: 1. The phosphorus of DNA in chloroplast appears to be transferred from inorganic phosphate, while in whole cell system from phosphate pool. 2. $^{32}P-phosphate$ content of acid soluble fraction in chloroplast as well as in whole cell system was more increased in the light than dark during the reaction. It was noted to be caused by the stimulation of sugar phosphate synthesis in the light. 3. It was confirmed that polyphosphate exists in chloroplast as well as whole cell. Acid insoluble polyphosphate content in whole cell system was significantly decreased during the reaction and the similar tendency was also observed in chloroplst. It is, therefore, considered that acid insoluble polyphosphate also play an most important role as a phosphate pool respectively in chloroplast and in cytoplasm. 4. Protein and lipid phosphorus in chloroplast as well as whole cell system were transferred from acid insoluble polyphosphate.

• Archives of Pharmacal Research
• /
• v.3 no.1
• /
• pp.7-12
• /
• 1980

Ampliciline was synthesized from 6-amino-pencillanic acid (6-APA) and D-.alpha. phenylglycine methyl ester by using amplicilin synthesizing enzyme from Peudomonas melanogenum (IAM 1655). The whole cell enzyme was immobilized by entrapping it in the polyacrylamide gel lattices. The polymer used in the enzyme entrapment was made from 150 mg per ml of acrylamide monomer and 8 mg per ml of N, N'-methylenebisacrylamide. About 200 mg/whole cell enzyme was mixed in the polymer for entrapment. The maximal activity retention after immobilization was 56%. The optimal pH values for the whole cell enzyme and the immobilized whole cell enzyme were 6.0 and 5.9, respectively. The optimal temperature for the enzyme activity were the same for both type of preparations. The enzyme stabilities against pH and heat increased for immobilized whole cell enzyme. Immobilized cell was more stable especially in the acidic condition while both type were found to be very suceptible to thermal inactivation at a temperature above 4.deg.C. The kinetic constants obtained from Lineweaver-Burk plot based on two substate reaction mechanism showed somewhat higher value for immobilized whole cell enzyme as compared to the whole cell enzyme : the Km value for 6-APA were 7.0 mM and 12.5 mM while Km values for phenylglycine methyl ester were 4.5 mM and 8.2 mM, respectively. Using the immobilized whole cell enzyme packed in a column reactor, the productivity of ampiciline was studied by varying the flow rate of substrate solution. At the space velocity, SV, 0.14 hr$^{-1}$ the conversion was 45%. Operational stability found in terms of half life was 30 hr at SV = 0.2 hr.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.6
• /
• pp.3515-3519
• /
• 2013

Background: Effects of whole cell type immunization on mice Ehrlich tumours were evaluated. Materials and Methods: After preliminary study, mice were divided two major groups; $1{\times}1000$ and $100{\times}1000$ live Ehrlich cell transferred major groups, each divided into four subgroups (n: 10). Study groups were immunized with Ehrlich cell lysates in 0, 3, 7, $14^{th}$ days and after 30 days of last immunization, live Ehrlich cells were transferred. Mice were observed for six months and evaluated for total and cancer free days. Results: Out of $100{\times}1000$ cell transferred solid type study group, all study group mean and tumour free periods were statistically longer than control groups. All $1{\times}1000$ Ehrlich cell transferred study groups survived significantly longer than $100{\times}1000$ Ehrlich cell transferred groups. Conclusions: Ehrlich mice tumours were prevented and survival prolonged with whole cell type immunization. Effects are related to the number of transferred tumor cells.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.5
• /
• pp.786-790
• /
• 2006

Cellular components of Lactococcus lactis ssp. lactis (heat-killed whole cells, cytoplasm, and cell walls) were tested for their in vivo immunopotentiating activity. Peritoneal macrophages from mice orally administered with heat-killed whole cells exhibited significantly greater phagocytic activity than the groups administered with cell-wall fraction or cytoplasm fraction. The cytotoxicity of natural-killer cells was the highest in the group administered with whole cells, and the production of cytokines ($IFN-\gamma$, IL-2, and IL-12) in spleen cells was significantly higher, when cellular components were injected, and it tended to be higher in the cell-wall and cytoplasm groups than in the whole-cell group. Interestingly, the cytokine production of Peyer's patch cells was high, when cytoplasm fractions were administered. These results demonstrate that whole cells and cytoplasm and cell-wall fractions of L. lactis ssp. lactis have immunopotentiating activities, which are related to the stimulation of Peyer's patches.

• JSTS:Journal of Semiconductor Technology and Science
• /
• v.5 no.1
• /
• pp.1-10
• /
• 2005

This paper presents the development of a microsystem for whole blood purification and electrophysiological analysis of the purified cells. Magnetophoresis using continuous diamagnetic capture (DMC) was utilized for whole cell purification and electrical impedance spectroscopy (EIS) was utilized for electrophysiological analysis of the purified cells. The system was developed on silicon and plastic substrates utilizing conventional microfabrication technologies and plastic microfabrication technologies. Using the magnetophoretic microseparator, white blood cells were purified from a sample of whole blood. The experimental results of the DMC microseparator show that 89.7% of the red blood cells (RBCs) and 72.7% of the white blood cells (WBCs) could be continuously separated out from a whole blood using an external magnetic flux of 0.2 T. EIS was used as a downstream whole cell analysis tool to study the electrophysiological characteristics of purified cells. In this work, primary cultured bovine chromaffin cells and human red blood cells were characterized using EIS. Further analysis capabilities of the EIS were demonstrated by successfully obtaining unique impedance signatures for chromaffin cells based on the whole cell ion channel activity.

• Korean Journal of Microbiology
• /
• v.24 no.3
• /
• pp.276-287
• /
• 1986

Chlorella ellipsoidea were cultured in the media containing rifampicin for 7 days. Aliquot cells were taken out after the inoculation and at intervals during cultivation and growth rate of Chlorella cells was measured. In order to investigate the effect of rifampicin on the nucleic acid synthesis, nucleic acid and RNA polymerase were extracted from chloroplast isolated from these cells, and the contents of nucleic acid and activity of enzyme were measured to compared with those of the control. The inhibitory concentration of rifampicin on growth was 80 ppm. The DNA contents in chloroplasts isolated were decreased 60% to compared with control, whole cells were markedly decreased 70% by rifampicin. The contents of base in the RNA were decreased 46% by rifampicin in shole cell, and 77% of base contents were decreased in chloroplast. Rifampicin also inhibited the activity of RNA polymerase, therefore whole cell was decreased 10% of activity and chloroplasts were decreased 42% of activity.

• Korean Journal of Veterinary Research
• /
• v.26 no.2
• /
• pp.265-276
• /
• 1986

The bovine serum factor influencing the growth of swine testicle (ST) cell and the END effect of hog cholera SN test was studied. Throughout the experimental studies. following results were obtained and summarized. 1. Bovine whole serum of 16(76.2%) and 4(19.0%) samples out of 21 have shown a positive ST cell growth and the END effect, respectively. However, all of 21(100%) and 8(38.1%) samples out of 21 serum supernatant fractions, prepared from the bovine whole serum, have shown positive ST cell growth and END effect, respectively. 2. In the SDS-polyacrylamide gel electrophoretic analysis of the bovine whole serum and the supernatant fractions, ST cell growth inhibiting factor was proved present in globulin fraction and in whole gel plate as a diffusible component. 3. The END ineffective component present in the whole serum and its supernatant fraction was proved to be BVDV neutralizing antibody. 4. The difference of osmolarity, optical density, pH, degree of precipitant formation following heat cold treatment, A/G ratio as we11 as electrophoretic pattern and NDV SN index of the samples were not correlated to the degree of 57 cell growth and to the END effectiveness.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.7
• /
• pp.1206-1215
• /
• 2016

Ginsenosides are the major active ingredients in ginseng used for human therapeutic plant medicines. One of the most well-known probiotic bacteria among the various strains on the functional food market is Lactobacillus rhamnosus GG. Biocatalytic methods using probiotic enzymes for producing deglycosylated ginsenosides such as Rd have a growing significance in the functional food industry. The addition of 2% cellobiose (w/v) to glucose-free de Man-Rogosa-Sharpe broths notably induced β-glucosidase production from L. rhamnosus GG. Enzyme production and activity were optimized at a pH, temperature, and cellobiose concentration of 6.0, 40℃, and 2% (w/v), respectively. Under these controlled conditions, β-glucosidase production in L. rhamnosus GG was enhanced by 25-fold. Additionally, whole-cell homogenates showed the highest β-glucosidase activity when compared with disrupted cell suspensions; the cell disruption step significantly decreased the β-glucosidase activity. Based on the optimized enzyme conditions, whole-cell L. rhamnosus GG was successfully used to convert ginsenoside Rb1 into Rd.