• Journal of Microbiology and Biotechnology
• /
• 제28권12호
• /
• pp.2009-2018
• /
• 2018

Leuconostoc mesenteroides can be used to produce mannitol by fermentation, but the mannitol productivity is not high. Therefore, in this study we modified the chromosome of Leuconostoc mesenteroides by genetic methods to obtain high-yield strains for mannitol production. In this study, gene knock-out strains and gene knock-in strains were constructed by a two-step homologous recombination method. The mannitol productivity of the pat gene (which encodes phosphate acetyltransferase) deletion strain (${\Delta}pat::amy$), the fk gene (which encodes fructokinase) deletion strain (${\Delta}fk::amy$) and the stpk gene (which encodes serine-threonine protein kinase) deletion strain (${\Delta}stpk::amy$) were all increased compared to the wild type, and the productivity of mannitol for each strain was 84.8%, 83.5% and 84.1%, respectively. The mannitol productivity of the mdh gene (which encodes mannitol dehydrogenase) knock-in strains (${\Delta}pat::mdh$, ${\Delta}fk::mdh$ and ${\Delta}stpk::mdh$) was increased to a higher level than that of the single-gene deletion strains, and the productivity of mannitol for each was 96.5%, 88% and 93.2%, respectively. The multi-mutant strain ${\Delta}dts{\Delta}ldh{\Delta}pat::mdh{\Delta}stpk::mdh{\Delta}fk::mdh$ had mannitol productivity of 97.3%. This work shows that multi-gene knock-out and gene knock-in strains have the greatest impact on mannitol production, with mannitol productivity of 97.3% and an increase of 24.7% over wild type. This study used the methods of gene knock-out and gene knock-in to genetically modify the chromosome of Leuconostoc mesenteroides. It is of great significance that we increased the ability of Leuconostoc mesenteroides to produce mannitol and revealed its broad development prospects.

• 오토저널
• /
• 제15권1호
• /
• pp.28-36
• /
• 1993

Present-day there are two of theories which have considerable scientific support to explain the knock phenomenon in S.I. engine, the detonation theory and the autoignition theory. But they still have some problems to explain effects of knock parameters, i.e.. compression ratio, spark timing, mixture quality, engine speed, ect, on knocking process in S.I. engine. Accordingly, it is essential to find out whish is more adequate theory of two and to develop the method of analyzing knock phenomenon, that is the aim of this paper. The Authors develop the method of predicting transient temperature and pressure at the end-gas zone during the combustion period and analyze knocking process by this method based on the knock theories. The caluculated values based on the autoignition theory show reasonablly correct relations between knock parameters and knock process but there is no evidence of knock occurred by detonation theory through the calculation according to the all parameters. The authors find out that the autoignition theory is more adequate than detonation theory to analyze knocking process in S.I. engine.

• 한국자동차공학회논문집
• /
• 제5권3호
• /
• pp.202-208
• /
• 1997

The knocking is one of major parameters to improve engine performance in a spark ignition engine. Many researches have been carried out to identify them using cylinder pressure, vibration signal and so on. In the present study, measurement and analysis was conducted to set up the criteria of knock occurrence by using microphone signal. Cylinder pressure was measured for the reference signal of knocking. It has been observed that resonance frequencies of pressure wave are nearly independent of engine operating conditions such as engine speed, air fuel ratio, load and octane number of fuel within to limited experimental conditions. SDBP(sum of different band-pass data) method using resonance frequency of knock was proposed for estimating knock intensity. SDBP method is superior to identify knock occurrence and its intensity in case of sound pressure measurement.

• Asian-Australasian Journal of Animal Sciences
• /
• 제33권6호
• /
• pp.1023-1033
• /
• 2020

Objective: The efficiency of the knock-in process is very important to successful gene editing in domestic animals. Recently, it was reported that transient loosening of the nucleosomal folding of transcriptionally inactive chromatin might have the potential to enhance homologous recombination efficiency. The objective of this study was to determine whether histone deacetylases (HDAC) inhibitor and RAD51 recombinase (RAD51) expression were associated with increased knock-in efficiency on the β-casein (bCSN2) gene locus in mammary alveolar-large T antigen (MAC-T) cells using the transcription activator-like effector nucleases (TALEN) system. Methods: MAC-T cells were treated with HDAC inhibitors, valproic acid, trichostatin A, or sodium butyrate for 24 h, then transfected with a knock-in vector, RAD51 expression vector and TALEN to target the bCSN2 gene. After 3 days of transfection, the knock-in efficiency was confirmed by polymerase chain reaction and DNA sequencing of the target site. Results: The level of HDAC 2 protein in MAC-T cells was decreased by treatment with HDAC inhibitors. The knock-in efficiency in MAC-T cells treated with HDAC inhibitors was higher than in cells not treated with inhibitors. However, the length of the homologous arm of the knock-in vector made no difference in the knock-in efficiency. Furthermore, DNA sequencing confirmed that the precision of the knock-in was more efficient in MAC-T cells treated with sodium butyrate. Conclusion: These results indicate that chromatin modification by HDAC inhibition and RAD51 expression enhanced the homologous recombination efficiency on the bCSN2 gene locus in MAC-T cells.

• International Journal of Automotive Technology
• /
• 제6권2호
• /
• pp.133-139
• /
• 2005

In this study, a three-dimensional transient simulation with a knock model was performed to predict knock occurrence and autoignition site in a heavy-duty LPG engine. A FAE (Flame Area Evolutoin) premixed combustion model was applied to simulate flame propagation. The coefficient of the reduced kinetic model was adjusted to LPG fuel and used to simulate autoignition in the unburned gas region. Engine experiments using a single-cylinder research engine were performed to calibrate the reduced kinetic model and to verify the results of the modeling. A pressure transducer and a head-gasket type ion-probe circuit board were installed in order to detect knock occurrences, flame arrival angles, and autoignition sites. Knock occurrence and position were compared for different piston bowl shapes. The simulation concurred with engine experimental data regarding the cylinder pressure, flame arrival angle, knock occurrence, and autoignition site. Furthermore, it provided much information about in-cylinder phenomena and solutions that might help reducing the knocking tendency. The knock simulation model presented in this paper can be used for a development tool of engine design.

• 한국자동차공학회논문집
• /
• 제1권1호
• /
• pp.22-31
• /
• 1993

Spark-ignition engine knock is affected by engine operating conditions such as engine speed, spark timing and intake air temperature. In this study the effect of intake air temperature on knock characteristics was studied experimentally using a 4-cylinder carburetor spark-ignition engine. The cylinder pressure data at 2000rpm were taken for intake air temperature range of $30^{\circ}C$ to $80^{\circ}C$ with $10^{\circ}C$ interval. And 80 consecutive cycles were taken at each experimental condition. As the same spark timing, as the intake air temperature increased by $50^{\circ}C$, the mean knock intensity increased about 20kPa. This effect corresponds to that of spark timing advance of 3 crank angle degrees.

• 한국자동차공학회논문집
• /
• 제10권5호
• /
• pp.107-113
• /
• 2002

LPG has been well known as a clean alternative fuel for vehicles. Recently, several LPG engines for heavy duty vehicles have been developed, which can replace some diesel engines that are one of the main sources for air pollution in the urban area. Because cylinder bore of heavy duty LPG engine is larger than that of gasoline, the study of knock characteristics of LPG engine are needed. In this study, the knock characteristics were investigated with various engine speed, air excess ratios and LPG fuel compositions. Experimental results indicated that the Knock occurrence probability decreases with increasing engine speed and propane fraction of fuel. The Knock occurrence probability is highest at excess air ratio of 1 and decreases as the mixture strength became leaner.

• 한국연소학회:학술대회논문집
• /
• 한국연소학회 2000년도 제21회 KOSCO SYMPOSIUM 논문집
• /
• pp.17-23
• /
• 2000

Knock in SI engines causes physical damage to the piston and combustion chamber and lowers the thermal efficiency. The increase in compression ratio which can improve the thermal efficiency and engine performance has been limited by engine knock. So the need of making clear the knocking phenomenon has increased. This paper reviews the methods of knock detection, characterization and prediction of knock with the reduced chemical kinetic modeling.

• 한국연소학회지
• /
• 제5권2호
• /
• pp.29-35
• /
• 2000

Knock in SI engines causes physical damage to the piston and combustion chamber and lowers the thermal efficiency. The increase in compression ratio which can improve the thermal efficiency and engine performance has been limited by engine knock. So the need of making clear the knocking phenomenon has increased. This paper reviews the methods of knock detection, characterization and prediction of knock with the reduced chemical kinetic modeling.

• 대한기계학회논문집A
• /
• 제21권2호
• /
• pp.223-231
• /
• 1997

A variety of approaches have been investigated for the application of spark-ignition engine knock control. The control method implemented, here as "Fuzzy Control", has the advantage of not requiring the knowledge of a mathematical model of the controlled object and is more robust and flexible than conventional approaches. Knock control in this study is performed using vibration signal which is measured with accelerometer attached to the cylinder block of a 1498cc four-cylinder spark-ignition engine. The experimental results obtained with this method are compared with those obtained with a knock interval controller and with those of a conventional controller. Those results illustrate better performance in torque than knock interval controller and conventional controller.ontroller.