• Title/Summary/Keyword: buffer time

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Physico-chemical characteristics and ${\beta}-galactosidase$ activity of Lactobacillus plantarum from kimchi (김치에서 분리한 유산균 Lactobacillus plantarum의 이화학적 특성 및 ${\beta}-galactosidase$ 활성)

  • Kang, Mi-Seon;Rhee, Young-Hwan
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.54-59
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    • 1996
  • Three strains of inhibitory lactic acid bacteria (No. 49, No. 61, No. 75) against Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escheirchia coli(ATCC33694) and Bacillus subtilis(ATCC6633) were isolated from kimchi, and then, identified to be Lactobacillus plantarum after examinations of their biological and physiological characteristics. To investigate a possible application of these three lactobacilli in milk fermentation industry, we made yogurts and then evaluated their ${\beta}-galactosidase$ activities at various; incubation time(from 24 hrs to 72 hrs). The result of experiment was that ${\beta}-galactosidase$ activities were reached maximum at 48 hrs and that reduced gradually with the lapse of time. And the ${\beta}-galactosidase$ activity of lactobacilli, and their viable cell counts at $37^{\circ}C$ for 2 hrs under various pH conditions were investigated. ${\beta}-galactosidase$ activities of 3 strains were reduced 50% at pH 3.5, but there were no remaining activities at pH 2.5, and pH 1.5, respectively. The frequency of the survival cell of lactobacilli in yogurt were $0.12{\sim}0.75%$ at pH 2.5, $$6.3{\times}10^{-5}{\sim}2.7{\times}10^{-3}% at pH 1.5, respectively, but there was no significant difference at pH 3.5. The values of original pH, titratable acidity as lactic acid, viscosity, and viable cells of yogurts were $4.08{\sim}4.30,\;1.05{\sim}1.25%,\;1,818{\sim}2,124\;cps\;and\;7.3{\times}10^8{\sim}3.0{\times}10^9\;cfu/m{\ell}$, respectively. To estimate buffer capacity of yogurt, the volume of 1.0 N HCl to 2 unit below original pH of yogurt($100\;m{\ell}$) was $11.98{\sim}13.02\;m{\ell}$ and the volume of 1.0N NaOH to 4 unit above original pH of yogurt($100\;m{\ell}$) was $10.82{\sim}12.86\;m{\ell}.

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Detection of Megalocytivirus in shellfish using PCR with various DNA extraction methods (다양한 PCR용 DNA 추출법에 의한 패류 내 Megalocytivirus의 검출)

  • Kim, Jin-Woo;Cho, Mi-Young;Jin, Ji-Woong;Kim, Ki-Hong;Jeong, Hyun-Do;Kim, Kwang-Il
    • Journal of fish pathology
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    • v.24 no.2
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    • pp.65-73
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    • 2011
  • In analysis of DNA viruses from the contaminated shellfish using PCR, preparation method of template DNA is an important factor to get enough copy number of viruses. In this study, we evaluated the efficiency of PCR template of Megalocytivirus (sT50mg-D) DNA obtained from 50 mg digestive gland homogenate of oyster using commercial method, and compared with that obtained from 5 g of the same tissues (T5g-D) after PEG precipitation procedures of virus. Both templates DNA suspended in the same volume of distilled water showed positive results by primary PCR with 35 cycles, and the presence of Megalocytivirus was confirmed in oysters collected from cultured farms in Korea. Moreover, PCR with sT50mg-D allowed us to discriminate the contaminated oyster individually, that can not be done in PCR with T5g-D prepared from the mixture of three different individual oyster to get 5 g digestive gland homogenate. In quantitative analysis with real time PCR, Megalocytivirus concentrations in 50 ${\mu}l$ templates prepared using 0.5~50 mg of one positive sample were appeared in the range 6.14E+00~1.2E+02/${\mu}l$. We were not able to get positive result using template DNA contained less than 6.14E+00 copies. Consequently, 2-step PCR performed with DNA extracts from oyster homogenate of small amount (sT50mg-D) i) was enough to detect the contaminated Megalocytivirus in shellfish, ii) allowed us to do the analysis for individual shellfish rather than mixture of several shellfish and iii) showed the presence of Megalocytivirus in oyster from Korea.

Changes in the Stability Properties of Methylcellulose Emulsions as Affected by Competitive Adsorption Between Methylcellulose and Tween 20 (메칠셀룰로오스/Tween 20 경쟁 흡착이 메칠셀룰로오스 유화액의 안정도 특성에 미치는 영향)

  • Hong, Soon-Taek
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1278-1286
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    • 2008
  • The effect of Tween 20 addition on changes in the stability of methylcellulose (MC) emulsions (1 wt% MC, 10 wt% n-tetradecane, 20 mM bis-tris buffer, pH 7) was investigated by creaming stability and orthokinetic stability measurements. In the case of MC emulsions containing varying amounts of oil (1$\sim$30 wt%) and no Tween 20 added, creaming stability, judged by mean migration velocity of fat globules ($V_m$), was found to depend on droplet size: the larger the droplet size, the worse the stability [$V_m$: 0.326 $\mu$m $min^{-1}$ ($d_{32}$: 0.32 $\mu$m) ${\rightarrow}V_m$: 0.551 $\mu$m $min^{-1}$ ($d_{32}$: 0.53 $\mu$m)]. With Tween 20, creaming stability was found to be worse than the one without Tween 20, except for MC emulsion containing 0.2 wt% Tween 20. In addition, cream stability was the lowest with the lowest concentration of Tween 20 and a tendency to recover with increasing Tween 20 concentration [$V_m$: 0.598 $\mu$m $min^{-1}$ (0.01 wt%)${\rightarrow}V_m$: 0.389 $\mu$m $min^{-1}$ (0.2 wt%)] was found. From viscosity measurement for aqueous bulk phase of MC emulsions, such a change in the creaming stability was found to coincide well with the results of viscosity measurement. Therefore, it was reasonable to say that creaming stability of MC emulsions containing Tween 20 depended on MC concentration in aqueous bulk phase, which was in turn varied by competitive adsorption between MC and Tween 20 at the oil droplet surface. In case of orthokinetic stability, judged by destabilization time ($t_d$), it was found that the addition of Tween 20 resulted in lowered stability with more pronounce tendency at higher concentrations [$t_d$: 160 min (0.03 wt%)${\rightarrow}t_d$: 100 min (0.2 wt%)]. Moreover, combined with previous results, the orthokinetic stability of MC emulsions containing Tween 20 was found to be exponentially proportional to MC load. In conclusion, competitive adsorption between MC and Tween 20 may affect the stability properties of MC emulsion to varying extents, depending on the concentration of Tween 20.

Interaction of Sodium Selenite on Neurotoxicity Induced by Methylmercuric Chloride (유기수은의 신경독성에 대한 셀레늄의 보상작용)

  • Park, J.S.;Lee, H.M.;Chung, Y.;Shin, D.C.;Roh, J.H.;Moon, Y.H.
    • Journal of Preventive Medicine and Public Health
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    • v.25 no.1 s.37
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    • pp.13-25
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    • 1992
  • This study was conducted to investigate the mechanism of protective effect by sodium selenite in methylmercuric chloride neurotoxicity, increasing intracellular $Ca^{2+}$concentration of the neuron. Methylmercuric chloride of 3mg/kg of body weight was administered simultaneously with sodium selenite of 5mg/kg and pretreatment of sodium selenite via intraperitoneal injection to rats. Also, effect of methylmercuric chloride($25{\mu}M,\;50{\mu}M,\;100{\mu}M$) and sodium selenite($200{\mu}M$) on free intrasynaptosomal $Ca^{2+}$ concentration were studied using the fluorescent $Ca^{2+}$ indicator fura -2 in vitro. After the treatment, at 6, 24, and 48 hours later, mercury in the cerebral cortex, liver and kidney tissues, succlnic dehydrogenase activities, adenosin-5'-triphosphate concentration, acetylcholinesterase activities, and intracellular $Ca^{2+}$ concentration in the cerebral cortex were determined in vivo. Cerebral synaptosomes of rats were incubated with methylmercuric chloride and sodium selenite in Hepes buffer for 10 minutes and free intrasynaptosomal $Ca^{2+}$ concentration were measured with fura-2 in vitro. The results were summarized as follows ; 1. The combined administration of $CH_3HgCl$ and $Na_2SeO_3$ and pretreatment of $Na_2SeO_3$ according to time significantly more increased in the cerebral cortex and decreased in the liver, kidney mercury concentrations compared to the administration of $CH_3HgCl$ only. 2. The combined administration of $CH_3HgCl$ and $Na_2SeO_3$ and pretreatment of $Na_2SeO_3$ increased more succinic dehydrogenase and acetylcholinesterase activities compared to the administration of $CH_3HgCl$ only. Particularly pretreatment of $Na_2SeO_3$ significantly more compared to the administration of $CH_3HgCl$ only. The concentration of adenosine-5'-triphosphate in $Na_2SeO_3$ treatment groups revealed a favourable effect compared to the administration of $CH_3HgCl$ only. 3. Intracellular $Ca^{2+}$ concentration in administration of $CH_3HgCl$ only was increased significantly more than control group in all test hours but was increased significantly more at 48 hours only after treatment in combined administration of $CH_3HgCl$ and $Na_2SeO_3$ and pretreatment of $Na_2SeO_3$ according to time interval more decreased significantly intracellular $Ca^{2+}$ concentration compared to the administration of $CH_3HgCl$ only. 4. Free intrasynaptosomal $Ca^{2+}$ concentration in the combined administration of $CH_3HgCl$ and $Na_2SeO_3$ was decreased ($24%{\sim}40%$) significantly more than the administration of $CH_3HgCl$ only. From the above results, the specific dosage of $Na_2SeO_3$ decreased increment of intracellular $Ca^{2+}$ concentration induced by administration of $CH_3HgCl$. These findings suggest the protective mechanism of $Na_2SeO_3$ on the neurotoxicity of $CH_3HgCl$.

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Protoplast Formation, Regeneration and Reversion in Pleurotus ostreatus and P. sajor-caju (느타리버섯과 여름느타리버섯의 원형질체(原形質體) 나출(裸出)과 재생(再生))

  • Go, Seung-Joo;Shin, Gwan-Chull;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.169-177
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    • 1985
  • The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.

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Estimation of the CY Area Required for Each Container Handling System in Mokpo New Port (목표 신항만의 터미널 운영시스템에 따른 CY 소요면적 산정에 관한 연구)

  • Keum, J.S.
    • Journal of Korean Port Research
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    • v.12 no.1
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    • pp.35-46
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    • 1998
  • The CY can be said to function in various respect as a buffer zone between the maritime and overland inflow-outflow of container. The amount of storage area needed requires a very critical appraisal at pre-operational stage. A container terminal should be designed to handle and store containers in the most efficient and economic way possible. In order to achieve this aim it is necessary to figure out or forecast numbers and types of containers to be handled, CY area required, and internal handling systems to be adopted. This paper aims to calculate the CY area required for each container handling system in Mokpo New Port. The CY area required are directly dependent on the equipment being used and the storage demand. And also the CY area required depends on the dwell time. Furthermore, containers need to be segregated by destination, weight, class, FCL(full container load), LCL(less than container load), direction of travel, and sometimes by type and often by shipping line or service. Thus the full use of a storage area is not always possible as major unbalances and fluctuations in these flow occuring all the time. The calculating CY area must therefore be taken into account in terms of these operational factors. For solving such problem, all these factors have been applied to estimation of CY area in Mokpo New Port. The CY area required in Mokpo New Port was summarized in the conclusion section.

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Studies on Xylooligosaccharide Analysis Method Standardization using HPLC-UVD in Health Functional Food (건강기능식품에서 HPLC-UVD를 이용한 자일로올리고당 시험법의 표준화 연구)

  • Se-Yun Lee;Hee-Sun Jeong;Kyu-Heon Kim;Mi-Young Lee;Jung-Ho Choi;Jeong-Sun Ahn;Kwang-Il Kwon;Hye-Young Lee
    • Journal of Food Hygiene and Safety
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    • v.39 no.2
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    • pp.72-82
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    • 2024
  • This study aimed to develop a scientifically and systematically standardized xylooligosaccharide analytical method that can be applied to products with various formulations. The analysis method was conducted using HPLC with Cadenza C18 column, involving pre-column derivatization with 1-phenyl-3-methyl-5-pyrazoline (PMP) and UV detection at 254 nm. The xylooligosaccharide content was analyzed by converting xylooligosaccharide into xylose through acid hydrolysis. The pre-treated methods were compared and evaluated by varying sonication time, acid hydrolysis time, and concentration. Optimal equipment conditions were achieved with a mobile phase consisting of 20 mM potassium phosphate buffer (pH 6)-acetonitrile (78:22, v/v) through isocratic elution at a flow rate of 0.5 mL/min (254 nm). Furthermore, we validated the advanced standardized analysis method to support the suitability of the proposed analytical procedure such as specificity, linearity, detection limits (LOD), quantitative limits (LOQ), accuracy, and precision. The standardized analysis method is now in use for monitoring relevant health-functional food products available in the market. Our results have demonstrated that the standardized analysis method is expected to enhance the reliability of quality control for healthy functional foods containing xylooligosaccharide.

The Effect of the Histidine-Tryptophan-Ketoglutarate (HTK) Solution on Myocardial Protection in Isolated Rat Heart (흰쥐의 적출심장에서 HTK 용액의 심근보호 효과)

  • 송원영;장봉현;김규태
    • Journal of Chest Surgery
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    • v.37 no.8
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    • pp.632-643
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    • 2004
  • Background: The Histidine-Tryptophan-Ketoglutarate (HTK) solution has been shown to provide the excellent myocardial protection as a cardioplegia. The HTK solution has relatively low potassium as an arresting agent of myocardium, and low sodium content, and high. concentration of histidine biological buffer which confer a buffering capacity superior to that of blood.. Since HTK solution has an excellent myocardial protective ability, it is reported to protect myocardium from ischemia for a considerable time (120 minutes) with the single infusion of HTK solution as a cardioplegia. The purpose of this study is to evaluate the cardioprotective effect of HTK solution on myocardium when the ischemia is. exceeding 120 minutes at two different temperature (10 to 12$^{\circ}C$, 22 to 24$^{\circ}C$) using the Langendorff apparatus, Material and Method: Hearts from Sprague-Dawley rat, weighing 300 to 340 g, were perfused with Krebs-Henseleit solution at a perfusion pressure of 100 cm $H_2O$. After the stabilization, the heart rate, left ventricular developed pressure (LVDP), and coronary flow were measured. Single dose of HTK solution was infused into the ascending aorta of isolated rat heart and hearts were preserved at four different conditions. In group 1 (n=10), hearts were preserved at deep hypothermia (10∼12$^{\circ}C$) for 2 hours, in group 2 (n=10), hearts were preserved at moderate hypothermia (22∼24$^{\circ}C$) for 2 hours, in group 3 (n=10), hearts were preserved at deep hypothermia for 3 hours, and in group 4 (n=10), hearts were preserved at moderate hypothermia for 3 hours. After the completion of the preservation, the heart rate, left ventricular developed pressure, and coronary flow were measured at 15 minutes, 30 minutes, and 45 minutes after the initiation of reperfusion to assess the cardiac function. Biopsies were also done and mitochondrial scores were counted in two cases of each group for ultrastructural assessment. Result: The present study showed that the change of heart rate was not different between group 1 and group 2, and group 1 and group 3. The heart rate was significantly decreased at 15 minutes in group 4 compared to that of group 1 (p<0.05 by ANCOVA). The heart rate was recovered at 30 minutes and 45 minutes in group 4 with no significant difference compared to that of group 1. The decrease of LVDP was significant at 15 minutes, 30 minutes and 45 minutes in group 4 compared to that of group 1 (p < 0.001 by ANCOVA). Coronary flow was significantly decreased at 15 minutes, 30 minutes, and 45 minutes in group 4 compared to that of group 1 (p < 0.001 by ANCOVA). In ultrastructural assessment, the mean myocardial mitochondrial scores in group 1, group 2, group 3, and group 4 were 1.02$\pm$0.29, 1.52$\pm$0.26, 1.56$\pm$0.45, 2.22$\pm$0.44 respectively. Conclusion: The HTK solution provided excellent myocardial protection regardless of myocardial temperature for 2 hours. But, when ischemic time exceeded 2 hours, the myocardial hemodynamic function and ultrastructural changes were significantly deteriorated at moderate hypotherma (22∼ 24$^{\circ}C$). This indicates that it is recommended to decrease myocardial temperature when myocardial ischemic time exceeds 2 hours with single infusion of HTK solution as a cardioplegia.

Simultaneous Determination and Mornitoring of Aflatoxin and Ochratoxin A in Food (식품 중 아플라톡신과 오크라톡신 A의 동시분석법 개발 및 모니터링)

  • Park, Ji-Won;Yoo, Myung-Sang;Kuk, Ju-Hee;Ji, Young-Ae;Lee, Jin-Ha
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.75-82
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    • 2013
  • The simultaneous analysis and monitoring of aflatoxin $B_1$, $G_1$, $B_2$, $G_2$ and ochratoxin A in foods were carried out by HPLC with fluorescence detection. The samples were extracted with methanol/water mixture. The extract was centrifuged, diluted with phosphate buffer saline (PBS), filtered, and applied to an immunoaffinity column containing antibodies specific to both aflatoxins and ochratoxin A. After washing the column with PBS and water, the toxins were eluted from the column with methanol, and quantified by HPLC, with a run time of approximately 30 min. The recoveries for aflatoxin $B_1$, $G_1$, $B_2$, $G_2$ and ochratoxin A in foods were 78.4~101.5%, 73.3~102.1%, 81.7~106.7%, 67.0~104.6% and 78.7~120.8%, respectively. The limits of detection of aflatoxins and ochratoxin A ranged from 0.05 to $0.18{\mu}g/kg$. According to monitoring result with the established method, aflatoxin $B_1$ and ochratoxin A were found in 13 of 151 domestic commercial foods. The contamination levels were $0.32{\sim}1.80{\mu}g/kg$ for aflatoxin $B_1$ and $0.97{\mu}g/kg$ for ochratoxin A. Therefore, this study showed all commercial foods monitored were safe under the Korean standards for aflatoxins and ochratoxin A.

Studies on the Hydrolysis of Wood with Dilute Sulphuric Acid Solution and Trichoderma viride Cellulase (묽은황산(黃酸) 및 Cellulase에 의(依)한 목재당화(木材糖化)에 관(關)한 연구(硏究))

  • Chung, In-Pyo;Kim, Hong-Eun;Min, Du-Sik
    • Journal of Korean Society of Forest Science
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    • v.41 no.1
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    • pp.1-6
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    • 1979
  • 1. The study was conducted on the optimum condition of the treated substrate with dilute sulphuric acid solution and cellulase for saccharification. The wood (saw dust) of Alnus hirsuta Rupr. (10~15 years) was treated with 0.3%, 0.6%, 0.9%, 1.2%, 1.5%, $H_2SO_4$ solution at $1.5kg/cm^2$ for 15min., 30min., 45min., and 60min., followed by thermal treatment at $190^{\circ}C$ for 30min., and screening with 60 mesh sieve, after which to 0.5 grams of each sample was added 0.5ml cellulase solution, and 50ml 0.1M acetic acid buffer solution (pH 5.0), after incubating at $40^{\circ}C$ for 96hr. 2. The crude cellulase of Trichoderma viride Perx. ex. Fr. SANK 16374 was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfite. 3. Reducing sugar was determined by the 3.5-dinitrosalicylic acid (DNS) method. 4. The reducing sugar was increased with increase of the sulphuric acid concentration and saw dust was treated with 1.5% $H_2SO_4$ solution at $1.5kg/cm^2$ for 45min. showed the best saccharification (16.0%). The reducing sugar formation did not show statistically significant in 5% levels by thermal treatment time 45min. and 60min. 5. The substrate for cellulase which was treated with 0.9% $H_2SO_4$ solution at $1.5kg/cm^2$ for 60min. showed the best reducing sugar formation (23.6%). And did not show significant difference in 5% levels at 0.9%, 1.2%, and 1.5% $H_2SO_4$ solution.

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