• 환경생물
• /
• 제27권4호
• /
• pp.406-413
• /
• 2009

The enzyme invertase contributes to sugar unloading, pathogen defense, differentiation and development in plants. We cloned the complete cDNA of a soluble acid invertase from pea seedlings (Pisum sativum) via RT-PCR and the rapid amplification of the cDNA end (RACE) technique. The full-length cDNA of the soluble pea invertase comprised 2237 bp and contained a complete open reading frame encoding 647 amino acids. The deduced amino acid sequence showed high homology to soluble acid invertases from various plants. Northern blot analysis demonstrated the soluble acid invertase gene of P. sativum was strongly expressed in sink organs such as shoot tips and root tips, and induced by abscisic acid, gibberellic acid and jasmonic acid in shoots. Especially, gibberellic acid enhanced the gene expression of the soluble acid invertase in a time-dependent manner. This study presents that the gene expression patterns of a soluble acid invertase from pea are strongly consistent with the suggestion that individual invertase gene product has different functions in the growing plant.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
• /
• 제2권1호
• /
• pp.37-48
• /
• 1998

Effect of plant hormones on the leaf senescence of mung bean (Phaseolus radiatus) was investigated by measuring the changes of reducing sugar contents and invertase isozyme activities in detached leaves treated with NAA, $GA_3$ or BA. During dark-induced senescence, reducing sugar contents in the detached leaves increased temporarily at 4 d, thereafter decreased rapidly and reached minimum values within 7-14 d. The pattern of soluble acid invertase activity in the senescing leaves kept in the dark was similar to that of reducing sugar accumulation, whereas the activities of alkaline and extracellular invertases were not significantly changed during leaf senescence. Therefore, these results suggest that soluble acid invertase, but not alkaline and extracellular invertases, induces the accumulation of reducing sugar during leaf senescence of mung bean plants. Exogenous NAA application had little or no effect in the increase of soluble acid invertase activity during dark-induced senescence compared to the control. However, exogenous applications of $GA_3$ and BA led to the increase of soluble acid invertase activity in the senescing leaves. Particularly, BA application was very effective in enhancing the activity of soluble acid invertase as well as in delaying chlorophyll breakdown during dark-induced senescence. These results suggest, therefore, that BA regulates the activity of soluble acid invertase, which leads to the accumulation of reducing sugar, and the stability of photosynthetic apparatus to delay leaf senescence.

• 한국환경과학회지
• /
• 제2권1호
• /
• pp.37-48
• /
• 1993

Effect of plant hormones on the leaf senescence of mung bean (Phseoln radiatus) was investigated by measuring the changes of reducing sugar contents and invertase isozyme activities in detached leaves treated with NAA, $GA_3$ or BA. During dark-induced senescence, reducing sugar contents in the detached leaves increased temporarily at 4 6, thereafter decreased rapidly and reached minimum values within 7-14 6. The pattern of soluble acid invertase activity in the senescing leaves kept in the dark was similar to that of reducing sugar accumulation, whereas the activities of alkaline and extracellular invertases were not significantly changed during leaf senescence. Therefore, these results suggest that soluble acid invertase, but not alkaline and extracellular invertases, induces the accumulation of reducing sugar during leaf senescence of Rung bean plants. Exogenous NAA application had little or no effect In the increase of soluble acid invertase activity during dark-induced senescence compared to the control. However, exogenous applications of $GA_3$ and BA led to the increase of soluble acid invertase activity in the senescing leaves. Particularly, BA application was very effective In enhancing the activity of soluble acid invertase as well as in delaying chlorophyll breakdown during dark-induced senescence. These results suggest, therefore, that BA regulates the activity of soluble acid invertase, which leads to the accumulation of reducing sugar, and the stability of photosynthetic apparatus to delay leaf senescence.

• Journal of Plant Biology
• /
• 제35권1호
• /
• pp.91-95
• /
• 1992

저온처리($4^{\circ}C$)하에서 보리(Hordeum vulgare L. cv. Chalssal) 유식물 잎의 당함량과 sucrosephosphate synthase, sucrose synthease, invertase의 활성의 변화를 측정하고, 또한 acid invertase의 성질을 조사하였다. 저온처리 3일만에 보리유식물의 환원당과 설탕의 함량은 1.3배 및 2.4배로 각각 증가하였다. 그리고 sucrosephosphate synthase, sucrose synthase 및 alkaline invertase의 활성에는 별다른 변화가 없었으나 acid invertase의 활성은 크게 저하하였다. Ammonium sulfate 분획과 DEAE-Sephacel 컬럼으로 부분정제한 acid invertase에서, 설탕에 대한 Km은 9.5mM이고 최적 pH는 5.5, 최적온도는 $35^{\circ}C$로 측정되었다. 여러 가지 기질에 대한 특이성을 조사한 결과 본 효소는 ${\beta}-fructosidase로$ 추정할 수 있었으며, 효소의 분자량은 Sephadex G-200 컬럼 크로마토그래피로 산출하였을 때 63 Kd이었다.

• 한국미생물·생명공학회지
• /
• 제20권3호
• /
• pp.348-354
• /
• 1992

SUC2 유전자를 가진 재조합 Saccharomyces cerevisiae의 invertase 생산성을 향상시키기 위하여 균체 생육과 유전자 발현에 미치는 아미노산과 용존산소 농도의 영향을 연구하였다. 균체 생육과 invertase 발현에 적합한 leucine과 histidine의 최적 첨가량은 탄소원인 포도당에 대한 비율로서 나타내어 0.03g/g glucose와 0.04g/g glucose였다. 회분배양에서는 통기량이 적을수록 invertase 비활성이 증가하였다. 희석율 0.09h에서 용존산소농도를 조절한 연속배양에서는 요존산소농도가 감소함에 따라 유전자 발현이 잘되어 5 포화 이하일때 invertase 비활성이 급격히 증가하여 통기를 하지 않은 조건에서 125.54KU/g cell의 최고 비활성을 얻었다.

• Journal of Ginseng Research
• /
• 제14권1호
• /
• pp.21-26
• /
• 1990

The chemical composition and stabilities of the purified ginseng invertase were investigated. The purified enzyme was found to be a glycoprotein composed of 80.2% protein and 19.7% total sugar. The protein component of the enzyme was composed of acidic amino acid (9.3%), basic amino acid (48.9%), nonpolar amino acid (21.4%), polar amino acid (20.4%) and 6.1% S-containing amino acid. It showed especially high contents of histidine and serine. The enzyme was inactivated almost completely by the treatment with some proteases (papain, pepsin. trypsin, pancreatin and microbial alkaline pretense) and protein denatllrants (8M urea and 6M guanidine-HC1), bolt not with glyrosidase (${\alpha}$-amylase, ${\beta}$-amylase. glcoamylese and cellullase). btonosaccharides sllch as glilrose, fructose, galactose and mannose did not exert any influence on the enzyme activity. The activity of the enzyme was inhibited by Ag+, Mn2+, Hg2+, Zn2+ and Al3+, whereas Ca2+, Mg2+, Ba2+ and Fe3+ gave rather activating effects on the enzyme activity. The enzyme was relatively stable in the VH range of VH 6 and 8, and at the temperatures below 35$^{\circ}C$.

• 생명과학회지
• /
• 제15권3호
• /
• pp.499-504
• /
• 2005

본 연구는 몇 가지 생장억제제를 배지에 처리하였을 때 기내 배양한 나도풍란 유묘의 생장과 그들의 invertase활성 검정을 위하여 실시하였다. 나도풍란에 식물생장억제제 인 ancymidol과 paclobutrazol을 각각 배양용 배지에 첨가했을 때, 농도가 높을수록 엽장이 감소한 반면, 엽폭은 상대적으로 증가하였고, 근장은 짧았으나 근수 및 근직경은 증가하였다 Uniconazole 0.05 mg/L에서는 잎과 뿌리생장이 대조구보다 오히려 촉진되었다. 처리한 모든 종류에 있어서 그 농도가 높아짐에 따라 잎 모양은 원형으로 변하면서 작아졌다. Invertase 활성을 알아보기 위 한 soluble acid invertase 활성과 soluble alkaline invertase활성은 잎에서는 생장억제제 농도가 낮은 처리에서 높았고, 뿌리에서는 정반대의 경향이었다. 지상부 및 지하부의 전분함량은 잎에서는 invertase 활성이 높았던 저농도에서 높았으며, 뿌리에서는 invertase 활성이 높았던 고농도에서 높았다.

• 생명과학회지
• /
• 제25권9호
• /
• pp.1021-1026
• /
• 2015

Invertase (β-D-fructosfuranosidase, EC 3.2.1.26)는 설탕을 포도당과 과당으로 가수분해하는 반응을 촉매한다. 3종류의 invertases [액포(수용성 산), 세포질(수용성 알칼리) 및 세포벽 결합]가 식물에서 연구되어 왔다. 우리는 순차적인 ammonium sulfate 침전, 이온교환크로마토그래피, 흡수크로마토그래피, Green-19 친화크로마토그래피 과정을 통해 완두콩(Pisum sativum L.) 발아체로부터 중성 invertase의 세포막 연결 isoform을 430배 순수 분리하였다. 분리된 세포막과 결합 된insoluble invertase (IN-INV)는 최적 pH는 중성에서 알칼리 사이(pH 6.8-7.5)로 나타났다. 이 효소는 Tris 뿐만 아니라 Hg²⁺ and Cu²⁺와 같은 중금속에 의해 저해되었다. IN-INV 의 K_m과 V_max 값은 각각 12.95 mM과 2.98 U/min으로 측정되었다. IN-INV는 기질로써 과당뿐만 아니라 라피노오스와 반응하기 때문에 진정한 β-fructofuranosidase로 판명되었다. IN-INV의 분자량 20 kDa이었다. 위 결과로 볼 때 GA 영향으로 급속히 자라는 발아체에서 단백질이 분리되었는데 특징적으로 invertase였다.

• Journal of Plant Biology
• /
• 제38권3호
• /
• pp.251-258
• /
• 1995

The soluble acid invertase ($\beta$-D-fructofuranoside fructohydrolase, EC 3.2.1.26) was isolated and characterized from the hypocotyls of mung bean (Phaseolus radiatus L.). The enzyme was purified to apparent homogeneity by consecutive step using diethylaminoethyl (DEAE)-cellulose anion exchange, Concanavalin (Con) A affinity and Sephacryl S-300 chromatography. The overall purification was about 148-fold with a yield of about 15%. The finally purified enzyme exhibited a specific activity of about 139 $\mu$mol of glucose produced mg-1 protein min-1 at pH 5.0 and appeared to be a single protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and nondenaturing PAGE. The enzyme had the native molecular weight of 70 kD and subunit molecular weight of 70 kD as estimated by Sephadex G-200 chromatography and SDS-PAGE, respectively, suggesting that the enzyme was composed of a monomeric protein. On the other hand, the enzyme appeared to be a glycoprotein containing N-linked high mannose oligosaccharide chain on the basis of its ability to bind to the immobilized C on A. The enzyme had a Km for sucrose of 1.8 mM at pH 5.0 and maximum activity around pH 5.0. The enzyme showed highest enzyme activity with sucrose as substrate, but the activity was slightly measured with raffinose and cellobise. No activity was measured with maltose and lactose. These results indicate the soluble acid invertase is a $\beta$-fructofuranosidase.

• 한국식품영양과학회지
• /
• 제21권6호
• /
• pp.725-730
• /
• 1992

이담자 효모균 Rhodosporidium toruloides의 mating type A 세포에서 세포내 invertase를 정제하였다. 세포내 invertase는 배양 세포의 파쇄액을 산침전 시킨 후 그 상등액으로부터 DEAE-Sephadex A-50, SP-Sephadex C-50 column chromatography와 Sephadex G-200 gel fitration 등의 과정을 거쳐 polyacrylamide gel disc 전기영동상 단일 효소 단백질까지 정제되었다. 정제효소의 분자량은 gel filtration에 의하여 90,000이었고, SDS-PAGE상에서는 22,000 dalttons에서 단일 band를 보여 단일종의 subunit가 4개로 구성된 단백질로 추정된다.