• Journal of Microbiology and Biotechnology
• /
• v.20 no.12
• /
• pp.1653-1663
• /
• 2010

The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.4
• /
• pp.431-439
• /
• 2014

We report the construction of two Bacillus subtilis expression vectors, pBNS1/pBNS2. Both vectors are based on the strong promoter P43 and the ampicillin resistance gene expression cassette. Additionally, a fragment with the Shine-Dalgarno sequence and a multiple cloning site (BamHI, SalI, SacI, XhoI, PstI, SphI) were inserted. The coding region for the amyQ (encoding an amylase) signal peptide was fused to the promoter P43 of pBNS1 to construct the secreted expression vector pBNS2. The applicability of vectors was tested by first generating the expression vectors pBNS1-GFP/pBNS2-GFP and then detecting for green fluorescent protein gene expression. Next, the mannanase gene from B. pumilus Nsic-2 was fused to vector pBNS2 and we measured the mannanase activity in the supernatant. The mannanase total enzyme activity was 8.65 U/ml, which was 6 times higher than that of the parent strain. Our work provides a feasible way to achieve an effective transformation system for gene expression in B. subtilis and is the first report to achieve B. pumilus mannanase secretory expression in B. subtilis.

• The Journal of Korean Institute of Electromagnetic Engineering and Science
• /
• v.15 no.4
• /
• pp.414-420
• /
• 2004

Partial transmit sequence(PTS) scheme has been widely studied to reduce the peak-to-average power ratio(PAPR) of Orthogonal frequency-division multiplexing(OFDM) signal since it is flexible ad any spectral regrowth is not made. A necessity of the PTS scheme is to transmit the side information about the phase rotation factors for correct OFDM symbol recovery. In this paper, a new side information embedded PTS scheme using the reference symbols is proposed fur the PAPR reduction and the BER(bit error rate) performance is analyzed when the erroneous side information is received. In this proposed method, the information about the rotation factors is expressed by the phase of reference symbols. The proposed method maintains the same PAPR reduction performance as the conventional PTS scheme and OFDM symbols are correctly recovered by the side information to meet the required BER level, which is verified via the computer simulation. Also, it is shown that the proposed method can be easily realized and can be used for all kinds of digital modulation formats in the OFDM communication system.

• The KIPS Transactions:PartB
• /
• v.8B no.6
• /
• pp.699-704
• /
• 2001

ABSTRACT In this paper, a new hybrid modular recurrent TDNN (time-delay neural network)-HMM (hidden Markov model) architecture for speech recognition has been studied. In TDNN, the recognition rate could be increased if the signal window is extended. To obtain this effect in the neural network, a high-level memory generated through a feedback within the first hidden layer of the neural network unit has been used. To increase the ability to deal with the temporal structure of phonemic features, the input layer of the network has been divided into multiple states in time sequence and has feature detector for each states. To expand the network from small recognition task to the full speech recognition system, modular construction method has been also used. Furthermore, the neural network and HMM are integrated by feeding output vectors from the neural network to HMM, and a new parameter smoothing method which can be applied to this hybrid system has been suggested.

• The Journal of Korean Institute of Communications and Information Sciences
• /
• v.30 no.12C
• /
• pp.1181-1185
• /
• 2005

A combining of MIMO signal processing with OFDM is regarded as a promising solution of enhancing the performance of next generation wireless system. Therefore, in this paper, an OFDM-based wireless system employing layered space-time architecture is considered for a high-rate transmission. In the MIMO-OFDM system, we evaluate the PAR performance using the SLM approaches. The investigated SLM scheme for MIMO-OFDM signals selects the transmitted sequence with lowest average PAR over all transmitting antennas and retrieves the side information very accurately at the expense of a slight degradation of the PAR performance. The low probability of false side information can improve the overall detection performance of the MIMO-OFDM system with erroneous side information compared to the ordinary SLM approache, respectively. Also, we provide closed form of the average BER performance in MIMO-OFDM system using analytic approach.

• The Journal of Korean Institute of Communications and Information Sciences
• /
• v.21 no.9
• /
• pp.2433-2443
• /
• 1996

In this paper, a new technique is proposed for obtaining the error probabilities of the VSB(vestigial sideband modulation) signal in the presence of the cochannel interference and frequency-selective fading channel. For the receivers, a suboptimal matched filter receiver and the MLSE(maximum likelihood sequence estimation) receiver, which is known to be optimal on the fading channel, are considered. First, for the matched filter receiver, the distributions of the random variables, which determine the SER(symbol error rate) are obtained by decomposing the multi-path fading channel into Rayleigh distributed main path and Gaussian distributed remained path channels. the random variables mean the energy of the main path and subpath respecitively, and SER can be calculated from the distribution of them. Next, for the case of the MLSE receover, it is found that the random variables are expressed as a function of integrals. In order to obtain the distribution for the random variables, we expanded each element of integrals with the KL(Karhunen-Loeve) transformation. And it is derived that the distributions for the transformed random variables are given by a sum of chi-square distributions. Finally, we calculated the error rate derived formula on the two-ray fading channel, which is one of widely used models for the frequency-selective fading channel. From the numerical results, it is found that for the matched filer receiver, performance degradation is significant, while the performance degradation at the MLSE receiver is insignificant on the frequency-selective fading channel. However, in case of cochannel interference environment, the error rateis found to increase significantly both at the matched filter and at the MLSE receiver.

• The Journal of Korean Institute of Communications and Information Sciences
• /
• v.19 no.5
• /
• pp.805-814
• /
• 1994

In this paper, we introduce a new method which is available for analyzing the throughput of the packet radio network by using the auxiliary Markov transient matrix with a failure state and a success state. And we consider the effect of symbol error for the network state(X, R) consisted of the number of transmitting PRU X and receiving PRU R. We examine the packet radio network of a continuous time Markov chain model, and the direct sequence binary phase shift keying CDMA radio channel with hard decision Viterbi decoding and bit-by-bit changing spreading code. For the unslotted distributed multi-hop packet radio network, we assume that the packet error due to a symbol error of radio channel has Poisson process, and the time period of an error occurrence is exponentially distributed. Through the throughputs which are found as a function of radio channel parameters, such as the received signal to noise ratio and chips of spreading code per symbol, and of network parameters, such as the number of PRU and offered traffic rate, it is shown that this composite analysis enables us to combine the Markovian packet radio network model with a coded DS/BPSK CDMA radio channel.

• BMB Reports
• /
• v.33 no.6
• /
• pp.493-498
• /
• 2000

Aquifex pyrophilus, a hyperthermophilic bacterium, has a serine-type protease that is located at the cell wall fraction with a mature size of 43 kDa. Molecular cloning of the protease gene revealed that it has an ORF of 619 amino acids with homologous catalytic site of serine-type proteases [Choi, I.-G., Bang, W.-K., Kim, S.-H., Yu, G. Y., J. Biol. Chem. (1999), Vol. 274, pp. 881-888]. Constructs containing different regions of the protease gene, including a alanine-substituted mutant at the active site serine, were constructed, and the factors affecting the expression level of the cloned protease gene in E. coli were examined. The presence of the C-terminus hydrophobic region of the protease hindered over-expression in E. coli. Also, the proteolytic activity of the expressed protein appeared to toxic to E. coli. An inactive form that deleted both of the N-terminal signal sequence and the C-terminal polar residues was over-expressed in a soluble form, purified to homogeneity, and its thermostability examined. The purified protein showed three disulfide bonds and three free sulfhydryl group. The thermal denaturation temperature of the protein was measured around $90^{\circ}C$ using a differential scanning calorimeter and circular dichroism spectrometry. The disulfide bonds were hardly reduced in the presence of reducing agents, suggesting that these disulfide bonds were located inside of the protein surface.

• BMB Reports
• /
• v.40 no.1
• /
• pp.22-28
• /
• 2007

MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.

• Progress in Medical Physics
• /
• v.7 no.1
• /
• pp.37-52
• /
• 1996

Image-guided localized, water-suppressed in vivo $^1$H MR spectroscopic studies were performed on the patients with brain tumors, acute cerebral infarction and schizophrenia, and dogs. GE Signa 1.5 T whole-body MRI/MRS system using STEAM pulse sequence was used. Proton metabolite ratios relative to creatine (Cr) were obtained using a Marquart algorithm. In vivo $^1$H MR spectra in brain neoplastic tissues revealed the changes of signal intensities of N-acetylaspartate (NAA), choline (Cho) and lactate (Lac) resonances. The present results suggest that the observed metabolite alterations from localized, water-suppressed in vivo $^1$H MR spectroscopy can be useful as an index of brain tumors, cerebral infarction and schizophrenia, and provide good quality metabolic information of cerebral tissue in the field of thanato-chronology.