• 한국토양비료학회지
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• 제45권1호
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• pp.51-58
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• 2012

Salinity is one of the most relevant abiotic factor limiting crop yield and its net primary productivity. In addition, salinity induces an increased stress ethylene synthesis in plants which, in turn, exacerbate the responses to the stressor. Bacterial single or co-inoculation effect was tested using previously characterized plant growth promoting (PGP) bacteria Brevibacterium iodinum RS16 and Methylobacterium oryzae CBMB20 on maize and sorghum-sudan grass hybrid under different concentrations of NaCl. Non-inoculated maize and sorghum-sudangrass hybrid showed 33.4% and 20.0% reduction in seed germination under highest NaCl (150 mM) level tested. However, under the same NaCl concentration, co-inoculation with B. iodinum RS16 and M. oryzae CBMB20 PGP strains increased the seed germination in maize (16.7%) and sorghum-sudangrass hybrid (4.4%). In Gnotobiotic growth pouch experiments conducted for maize and sorghum-sudangrass hybrid, co-inoculation of PGP B. iodinum RS16 and M. oryzae CBMB20 mitigated the salinity stress and promoted root length by 22.9% and 29.7%, respectively. Thus the results of this study could help in development of potential bioinoculants that may be suitable for crop production under saline conditions.

• 한국미생물·생명공학회지
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• 제9권1호
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• pp.45-50
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• 1981

Adenine-guanine 동시영양요구성인 Brevibacterium ammoniagenes D-21530을 사용하여 5'-IMP을 생산할 경우, 5'-IMP생성에는 탄소원으로 fructose가 glucose보다 효율적이었다. 반면에 균 생육에는 glucose를 탄소원으로 사용하는 것이 좋았다. Glucose와 fructose를 혼합하여 사용할 때 균 생육을 적절히 조절하여 5'-IMP축적을 증가시켜 줄 수 있었으며 fructose와 glucose의 혼합당에 있는 fructose의 함유율을 20∼40%범위로 하였을 때 5'-IMP생성이 최대가 되었고 fructose 단독으로 사용할 때보다 약 40% 증가되었다. 또한 영양요구성물질인 adenine과 guanine의 첨가농도가 증가함에 따라 균생육이 촉진되나 150㎎/l 이상에서는 정상상태였고, 5'-IMP생성은 50㎎/l 까지는 첨가 농도에 따라 급격히 증가하나 그 이상에서는 감소하였다.

• 한국토양비료학회지
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• 제47권1호
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• pp.1-7
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• 2014

The objective of this study was to determine the effect of single and co-inoculation of plant growth promoting bacteria (PGPB) on early plant growth in Saemangeum reclaimed soil. Plant growth promoting Brevibacterium iodinum RS16 and Methylobacterium oryzae CBMB20 were inoculated on maize (Zea mays L.) and sorghum-sudangrass hybrid (Sorghum bicolor L.) grown in Saemangeum reclaimed soil. Single and co-inoculation of B. iodinum RS16 and M. oryzae CBMB20 increased plant height, dry biomass accumulation and macro-nutrient accumulation of maize and sorghum-sudangrass hybrid. M. oryzae CBMB20 treatment increased plant height in maize by 41.2% at 30 days after sowing (DAS), shoot dry weight and total dry weight compared to non-inoculated treatment. Macro-nutrient accumulation (N and P) in maize roots was significantly increased with co-inoculation treatment, K and Ca content was significantly increased at B. iodinum RS16 treatment compared to non-inoculated treatment. Macro-nutrient accumulation (P, K, Ca and Mg) in shoot was higher with M. oryzae CBMB20 treatment compared to non-inoculated treatment. In case of sorghum-sudangrass hybrid, co-inoculation treatment showed 33.7% increase in plant height compared to non-inoculated treatment at 30 DAS. M. oryzae CBMB20 treatment increased root dry weight and total dry weight, macro-nutrient accumulation in roots and N, Ca and Mg accumulation in shoot compared to non-inoculated treatment. P and K accumulation in shoot was significantly increased at co-inoculation treatment compared to non-inoculated treatment. This pot culture experiment demonstrated that single and co-inoculation of B. iodinum RS16 and M. oryzae CBMB20 increased the early growth and nutrient accumulation of maize and sorghum-sudangrass hybrid.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.819-824
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• 2001

The principal DNA modification systems of the amino-acid-producing bacteria Corynebacterium glutamicum AS019, Brevibacterium flavum BF4, and B. lactofermentum BL1 was investigated using two approaches; digestion of plasmid DNA isolated from these species TseI and Fnu4HI, and sequence analysis of the putative methyltransferase target sites following the derivatization of DNA using metabisulfite treatment. The C. glutamicum and B. flavum strains showed similar digestion patterns to the two enzymes, indicating that the target for cytidine methyltransferase recognizes 5'-GCSGC-3'(where S is either G or C). Mapping the methylated cytidine sites by bisulfite derivatization, followed by PCR amplification and sequencing, was only possible when the protocol included an additional step eliminating any underivatized DNA after PCR amplification, thereby indicating that the derivatization was not $100\%$ efficient. This may have been due to the high G0C content of this genus. It was confirmed that C. glutamicum AS019 and B. flavum BF4 methylated the cytidine in the $Gm^5CCGC$ sequences, yet there were no similar patterns of methylation in B. lactofermentum, which was consistent with the distinctive degradation pattern seen for the above enzymes. These findings demonstrate the successful application of a modified bisulfite derivatization method with the Corynebacterium species for determining methylation patterns, and showed that different species in the geneus contain distinctive restriction and modification systems.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.214-221
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• 1998

A Brevibacterium ammoniagenes gene coding for glucose/mannose-specific enzyme II ($EII^{Glc}$) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing an Escherichia coli mutation affecting a ptsG gene, and the complete DNA nucleotide sequence was determined. The cloned gene was identified to be a ptsG, which enables the E. coli transportment to use glucose more efficiently than mannose as the sole carbon source in an M9 minimal medium. The ptsG gene of B. ammoniagenes consists of an open reading frame of 1,983 nucleotides putatively encoding a polypeptide of 661 amino acid residues and a TAA stop codon. The deduced amino acid sequence of the B. ammoniagenes $EII^{Glc}$ shows, at $46\%$, the highest degree of sequence similarity with the Corynebacterium glutamicum EII specific for both glucose and mannose. In addition, the $EII^{Glc}$ shares approximately $30\%$ sequence similarities with sucrose-specific and ${\beta}$-glucoside-specific EIIs of the several bacteria belonging to the glucose-PTS class. The 161-amino-acid C-terminal sequence of $EII^{Glc}$ is also similar to that of E. coli enzyme $IIA^{Glc}$, specific for glucose ($EIIA^{Glc}$). The B. ammoniagenes $EII^{Glc}$ consists of three domains; a hydrophobic region (EIIC) and two hydrophilic regions (EIIA, EIIB). The arrangement of structural domains, IIBCA, of the $EII^{Glc}$ is identical to those of EIIs specific for sucrose or ${\beta}$-glucoside. While the domain IIA was removed from the B. ammoniagenes $EII^{Glc}$ the remaining domains IIBC were found to restore the glucose and mannose-utilizing capacity of E. coli mutant lacking $EII^{Glc}$ activity with $EIIA^{Glc}$ of the E. coli mutant. $EII^{Glc}$ contains a histidine residue and a cysteine residue which are putative phosphorylation sites for the protein.

• 한국미생물·생명공학회지
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• 제1권2호
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• pp.75-78
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• 1973

핵산관연물질중 정미성물질인 5'-inosinic acid을 직접발효법으로 생산함에 있어 배지조성의 검토를 본 실험에서 수행하였다. 유기질소원으로서는 yeast extract 1%을 사용함이 좋았고 biotin의 첨가량은 30 $\mu\textrm{g}$l였을때 IMP의 축적량이 가장 많았다. 유기질소원으로 Casamino acid를 쓸 경우 $Mn^{＋＋}$ 20$\mu\textrm{g}$1. $Zn^＋＋/10$\mu\textrm{g}$1, Thiamine HCl 10mg/1 및 Ca-D-pantothenate 5 mg/l를 첨가해주면 IMP의 축적량이 증가하였으며 최고 7.0mg/ml의 IMP가 발효배지중에 생성되었다.었다.

• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.188-193
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• 1995

A Brevibacterium flavum gene coding for glucose permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing the Escherichia coli ZSCl13 mutations affecting a ptsG gene with the B. flavum genomic library. From the E. coli clone grown as red colony on a MacConkey plate supplemented with glucose as an additional carbon source, a recombinant plasmid was isolated and named pBFT93. The plasmid pBFT93 was identified as carrying a 3.6-kb fragment of B. flavum chromosomal DNA which enables the E. coli transformant to use glucose or man nose as a sole carbon source in an M9 minimal medium. The non-metabolizable sugar analogues, 2-deoxy-D-glucose (2-DG) and methyl-$\alpha$-D-glucopyranoside (MeGlc) affected the growth of ZSCl13 cells carrying the plasmid pBFT93 on minimal medium supplemented with non-PTS carbohydrate, glycerol, as a sole cabon source, while the analogues did not repress the growth of ZSCl13 cells without pBFT93. It was also found that both $2-deoxy-D-[U-^{14}C]glucose{\;}and{\;}methyl-{\alpha}-D-[U-^{14}C]glucopyranoside$ could be effectively transported into ZSCl13 cells transformed with plasmid pBFT93. Several in vivo complementation studies suggested that the B. flavum DNA in pBFT93 encodes a glucose permease specific for glucose and mannose.

• 한국식품과학회지
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• 제4권2호
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• pp.72-76
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• 1972

2,2'-methylene bis(3,4,6-trichloroacetoxy benzene)을 모체로 한 7가지 hydroxyamine 유도체의 13가지 균주에 대한 항균성을 paper disk method와 tube dilution method에 의하여 얻은 결과는 다음과 같다. 1) hydroxyamine유도체 중에서 -OH기가 meta위치에 있는 유도체가 다른 유도체보다 항균력이 강하고, para위치에 -OH기가 있는 화합물은 전연 어느 균에도 항균력이 없으며, para위치에 있는 -OH기의 H대신에 $CH_{3-}$기가 치환된 유도체도 항균력이 별로 강하지가 않았다. 2) $-NH\;OH,\;-NH\;CH_2\;CH_2OH$ 및 $-N\;(CH_2\;CH_2\;OH)_2$화합물 중에서 -NH OH화합물이 가장 강하고 나머지 두 화합물은 몇 가지 균주에 대하여 약간의 항균력을 나타내었다. 3) 모든 합성화합물 중에서 -NH OH화합물의 항균력이 제일 강했으며 Brevibacterium ammoniagenes에 대한 M.I.C.는 $1.6{\mu}g/ml$이며, S. aureus와 Bacillus subtilis에 대한 M.I.C.는 $5{\mu}g/ml$이었다.

• 한국식품과학회지
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• 제31권1호
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• pp.224-230
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• 1999

$_L-Lysine$ 발효산업에 이용되고 있는 B. lactofermentum의 L-lysine 생합성은 succinylase 경로와 dehydrogenase 경로를 통하여 일어난다. 특히 lysine 생산 균주에 부가적으로 존재하는 dehydrogenase 경로는 lysine 생합성에 있어서 필수적인 경로로 작용하며 이때 meso-DAP-dehydrogenase (DDH)를 암호화하는 ddh gene이 관여한다. 그러므로 B. lactofermentum의 lysine 발효에 있어서 ddh gene의 over expression에 의한 lysine 생성량을 비교 조사하기 위하여, shuttle vector pEB1 및 pJC1으로 ddh gene을 삽입하여 재조합 plasmid pRK1 및 pRK31을 구축하였고 이를 B. lactofermentum으로 도입시켜 DDH 활성을 측정한 결과 pRK1을 함유한 균주는 대조균주보다 7배 정도, pRK31을 함유한 균주는 14배 정도 증가하였다. 또한 Shuttle vector를 함유한 대조균주와 재조합 plasmid를 함유한 균주간의 성장 비교에서는 서로 비슷한 수준을 나타내었으며 플라스크 배양에서 lysine 생성량의 비교 분석에서는 재조합 plasmid를 함유한 균주의 경우 48시간 이후부터 대조균주보다 lysine 생성량이 증가하기 시작하여 72시간때에는 최대치를 나타내었으며 그 이후는 오히려 감소하였다. 최대치를 나타낸 72시간 때의 lysine 생성량은 대조균주가 4.38g/L를 나타내었으며 pRK1 및 pRK31을 함유한 균주는 각각 5.34g/L 및 5.21 g/L이었다. 이상의 결과로 미루어 볼 때 B. lactofermentum내에서 ddh gene의 증폭에 의한 lysine 생성량은 pRK1 및 pRK31에서 대조균주보다 각각 20% 및 19% 증가하였다. 또한 발효조 배양에서의 lysine 생성량도 재조합 균주가 대조균주보다 23% 정도 증가를 나타내어 B. lactofermentum에서 ddh gene의 증폭으로 lysine 생성량이 증가하였음을 확인하였다.

• 한국식품과학회지
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• 제13권2호
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• pp.121-126
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• 1981

Brevibacterium ammoniagenes ATCC 6872를 변이하여 얻은 adenine-guanine 및 $\beta$-alanine 영양요구성(營養要求性)이고 5'-XMP를 다량 생산하는 변이주 D-1550-40을 사용하여 발효배지성분(醱酵培地成分)의 최고농도(最高濃度)와 각 성분간의 상호작용효과(相互作用效果)를 검토하였다. 5'-XMP 축적(蓄積)에 필요한 adenine 및 guanine의 최적(最適) 농도(濃度)는 각각 75mg/l이었으나 생육(生育)에 대한 최적농도(最適濃度)는 이 보다 높은 100mg/ml이었고 그 이상의-농도에서는 5'-XMP 축적에 심한 저해를 초래하였다. 이러한 현상은 biotin을 $100{\mu}g/l$ 이상 첨가하거나 casamino acid를 0.3% 이상 첨가함으로서 배제할 수 있었다. 5'-lMP 발효의 경우와 마찬가지로 무기인산염(無機燐酸鹽)과 마그네슘은 5'-XMP 축적에도 $1.0{\sim}1.5%$1.5%가 적당(適當)하였다. $MnSO_4$의 농도가 $0{\sim}5mg/l$의 범위안에서 증가함에 따라 균증식과 5'-XMP의 축적은 촉진되었으나 그 이상의 농도에서는 변화가 없었고 정상상태를 나타내었다. Adenine과 guanine 각 100 mg/l, $MnSO_4\{cdot}7H_2O5mg/l 및 biotin $100{\mu}g/lg$가 함유된 발효배지에서 배양한 결과 4일후 60.5mg/ml의 5'-XMP가 측적되었으며, 5'-XMP의 생성활성(生成活性)은 균체량(菌體量)에 비례하여 배양 2일 내지 3일에서 가장 높았다.