• 환경생물
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• 제27권4호
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• pp.406-413
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• 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
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• 제2권1호
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• pp.37-48
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• 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
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• 제2권1호
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• pp.37-48
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• 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
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• 제38권3호
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• pp.251-258
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• 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.

• 생명과학회지
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• 제15권3호
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• pp.499-504
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• 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
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• 제25권9호
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• pp.1021-1026
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• 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 Life Science
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• 제11권1호
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• pp.14-21
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• 2001

To investigate the changes of carbohydrate metabolism in the senescing leaves of Zea mays during dark-induced senescence, the changes in the contents of reducing sugar, sucrose and starch as well as the activities of sucrose synthase, three isozymes of invertase, and ${\alpha}$-amylase were measured. In the senescing leaves, the content of reducing sugars temporarily increased at 4 d and rapidly decreased thereafter, whereas sucrose contents gradually decreased thereafter, whereas sucrose contents gradually decreased until 3 d of senscence and significantly decreased thereafter. The activities of intracellular invertases such as soluble acid and alkaline formed gradually enhanced until 4 d of leaf senescence and significantly declined thereafter. The extracellular invertase activity showed no significant changes during leaf senescence. The deactivation of sucrose synthase was observed within 3 d of leaf senscence. On the other hand, the starch contents gradually declined during 2 d of leaf senescence, and showed a temporary increase at 3 d, which is similar to the pattern of sucrose synthase activity., These results imply that sucrose in the senescing leaves. The major enzymes which correlated to the breakdown of sucrose during dark-induced senescence were soluble acid and alkaline invertases, not sucrose and ABA accelerated leaf senescence by inducing the accumulation of reducing sugar. These result, therefore, that leaf senescence may be mediated by the temporary quantitative changes of reducing sugar induced by the activation of intracellualr inveertases.

• 원예과학기술지
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• 제34권6호
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• pp.898-911
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• 2016

Sugars play many important roles in plant metabolism and directly influence fruit quality. The effects of two edible coatings, 2% calcium chloride and 1% pullulan, on sugar metabolism in postharvest Huangguan pear fruit were investigated during cold storage. The respiration rate, sugar content and composition, sucrose metabolism enzyme activities [acid invertase (AIV), neutral invertase (NI), sucrose synthase (SS), and sucrose phosphate synthase (SPS)] and expression of the AIV gene were analyzed during storage at $0^{\circ}C$ for 210 days. Coating treatments slowed the decrease of sucrose and hexose, the respiration rate, and the activities of AIV, NI, SS and SPS, thus maintaining high total soluble solids (TSS) and titratable acid (TA) contents in the fruit. There were no significant differences in AIV expression or activity between the treated and control groups of fruits. Both of the coatings could inhibit the activities of sucrose-cleaving enzymes, thus slowing the decrease of sugar content and maintaining high fruit quality during cold storage.

• Journal of Plant Biology
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• 제35권4호
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• pp.333-338
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• 1992

다양한 발달단계에 있는 은행종자의 자성배우체, 자엽 및 유근에서 탄수화물의 함량과 여러 가수분해효소(아밀라제, 인버타제 및 인산가수분해효소)의 활성을 비교하였다. 종자의 초기발아시에 아밀라제의 활성이 크게 증가함에 따라 자성배우체의 전분이 현저한 감소를 보인 반면 수용성 당들은 증가하였다. 인버타제의 활성은 건조한 종자에서는 매우 낮았으며 바라된 자엽과 유근에서 점진적으로 증가하였다. 인산가수분해효소의 활성은 유근이 신장되는 단계에서 변화를 나타내었으며 산성-인산가수분해효소가 알카리성-인산가수분해효소보다 더 높은 활성을 보였다.

• Journal of Plant Biology
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• 제34권2호
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• pp.113-119
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• 1991

저온 처리($4^{\circ}C$)가 벼(Oryza sativa L. cv. Samjin) 유식물 잎의 환원당, 설탕, 녹말 및 fructose-2,6-bisphosphate(F-2,$6-P_2$)의 함량에 미치는 영향을 조사하였으며, 이와 관련있는 amylase, invertase 및 fructose-1,6-bisphosphatase(FB-Pase)의 활성변화를 중심으로 3일 동안 측정하였다. 3일 동안 저온처리한 벼 유식물에서의 환원당과 설탕의 함량은 증가하였고, 수용성 녹말과 비수용성 녹말의 함량은 감소하였다. 한편, amylase의 활성은 증가하였고 acid invertase의 활성을 감소하였으며, alkaline invertase의 활성은 변화가 없었다. 스트로마 및 시토졸 FBPase의 활성은 감소하였다. 이에 부합되는 것으로 F-2,$6-P_2$의 함량이 감소하였다. 그러므로, 저온처리로 환원당의 함량이 증가한 것은 녹말 분해능의 증가에 기인한 것으로 사료되며, 설탕의 함량이 증가한 것은 설탕 분해능의 저하와 F-2,$6-P_2$의 함량감소에 따른 시토졸 FBPase의 활성증가에 기인한 것으로 해석된다. 또한 저온에 의한 이러한 탄수화물 대사의 변화는 벼 유식물의 저온 손상을 일부나마 방지하려는 기작이라 해석된다.