粗糙脉孢菌(Neurospora crassa)木糖发酵的研究

研究了不同通氧条件和培养基初始pH等对粗糙脉孢菌(Neurospora crassa)AS3．1602木糖发酵的影响。结果表明,粗糙脉孢菌具有较强的发酵木糖产生乙醇及木糖醇的能力。通气量对木糖发酵有较大的影响。乙醇发酵适合在半好氧条件下进行,此时乙醇的转化率达到63．2％。木糖醇发酵适合在微好氧的条件下进行,转化率达到31．8％。木糖醇是在培养基中乙醇达到一定浓度后才开始积累。培养基的初始pH对木糖发酵产物有较大的影响,乙醇产生最适pH5．0,木糖醇产生最适pH4．0。在培养基pH为碱性条件时,木糖发酵受到很大的抑制。初始木糖浓度对产物乙醇及木糖醇的产率有很大的影响。葡萄糖的存在会抑制木糖的利用,对乙醇和木糖醇的产生也有很大的影响。     

Influence of temperature and pH on xylitol production from xylose by Debaryomyces hansenii

The production of xylitol from concentrated synthetic xylose solutions (S(o) = 130-135 g/L) by Debaryomyces hansenii was investigated at different pH and temperature values. At optimum starting pH (pH(o) = 5.5), T = 24 degrees C, and relatively low starting biomass levels (0.5-0.6 g(x)/L), 88% of xylose was utilized for xylitol production, the rest being preferentially fermented to ethanol (10%). Under these conditions, nearly 70% of initial carbon was recovered as xylitol, corresponding to final xylitol concentration of 91.9 g(P)/L, product yield on substrate of 0.81 g(P)/g(S), and maximum volumetric and specific productivities of 1.86 g(P)/L x h and 1.43 g(P)/g(x) x h, respectively. At higher and lower pH(o) values, respiration also became important, consuming up to 32% of xylose, while negligible amounts were utilized for cell growth (0.8-1.8%). The same approach extended to the effect of temperature on the metabolism of this yeast at pH(o) = 5.5 and higher biomass levels (1.4-3.0 g(x)/L) revealed that, at temperatures ranging from 32-37 degrees C, xylose was nearly completely consumed to produce xylitol, reaching a maximum volumetric productivity of 4.67 g(P)/L x h at 35 degrees C. Similarly, both respiration and ethanol fermentation became significant either at higher or at lower temperatures. Finally, to elucidate the kinetic mechanisms of both xylitol production and thermal inactivation of the system, the related thermodynamic parameters were estimated from the experimental data with the Arrhenius model: activation enthalpy and entropy were 57.7 kJ/mol and -0.152 kJ/mol x K for xylitol production and 187.3 kJ/mol and 0.054 kJ/mol x K for thermal inactivation, respectively.     

The behavior of key enzymes of xylose metabolism on the xylitol production by <Emphasis Type="Italic">Candida guilliermondii</Emphasis> grown in hemicellulosic hydrolysate

A variety of raw materials have been used in fermentation process. This study shows the use of rice straw hemicellulosic hydrolysate, as the only source of nutrient, to produce high added-value products. In the present work, the activity of the enzymes xylose reductase (XR); xylitol dehydrogenase (XD); and glucose-6-phosphate dehydrogenase (G6PD) during cultivation of Candida guilliermondii on rice straw hemicellulosic hydrolysate was measured and correlated with xylitol production under different pH values (around 4.5 and 7.5) and initial xylose concentration (around 30 and 70 g l(-1)). Independent of the pH value and xylose concentration evaluated, the title of XD remained constant. On the other hand, the volumetric activity of G6PD increased whereas the level of XR decreased when the initial xylose concentration was increased from 30 to 70 g l(-1). The highest values of xylitol productivity (Q (P) approximately 0.40 g l(-1)) and yield factor (Y (P/S) approximately 0.60 g g(-1)) were reached at highest G6PD/XR ratio and lowest XR/XD ratio. These results suggest that NADPH concentrations influence the formation of xylitol more than the activity ratios of the enzymes XR and XD. Thus, an optimal rate between G6PD and XR must be reached in order to optimize the xylitol production.     

Effect of temperature on the microaerophilic metabolism of Pachysolen tannophilus

Xylitol production by Pachysolen tannophilus from detoxified hemicellulose hydrolysate was investigated under microaerophilic conditions at temperature ranging from 20 to 40 degrees C. A carbon balance previously proposed to study the influence of pH was used in this work to evaluate the amounts of carbon source (xylose) utilised in competitive metabolic ways: reductive production of xylitol, ethanol fermentation and respiration. At pH = 5.5 more than 83% of xylose was reduced to xylitol at 25 < T < 30 degrees C, whereas respiration became the main process at low temperature (71.1% at 20 degrees C). At high temperature, on the other hand, all three processes took place at comparable rate, consuming at 40 degrees C nearly the same percentage of carbon source (33-35%). Finally, the maximum values of volumetric productivity calculated at variable temperature were used to estimate the main thermodynamic parameters of both xylitol production (Deltah* = 105.4 kJ mol(-1); Deltas* = -13.2 J mol(-1) K(-1)) and thermal deactivation (Deltah*(D) = 210.5 kJ mol(-1); Deltas*(D) = 3.63 J mol(-1) K(-1)).     

Production of xylitol by recombinant Saccharomyces cerevisiae containing xylose reductase gene in repeated fed-batch and cell-recycle fermentations

Xylitol is a well-known sugar substitute with low-calorie and anti-cariogenic characteristics. An effort of biological production of xylitol from xylose was made in repeated fed-batch and cell-recycle fermentations of recombinant Saccharomyces cerevisiae BJ3505/δXR harboring the xylose reductase gene from Pichia stipitis. Batch fermentation with 20 g/l xylose and 18 g/l glucose resulted in 9.52 g/l dry cell mass, 20.1 g/l xylitol concentration and approximately 100% conversion yield. Repeated fed-batch operation to remove 10% of culture broth and to supplement an equal volume of 200 g/l xylose was designed to improve xylitol production. In spite of a sudden drop of cell concentration, an increase in dry cell mass led to high accumulation of xylitol at 48.7 g/l. To overcome loss of xylitol-producing biocatalysts in repeated fed-batch fermentation, cell-recycle equipment of hollow fiber membrane was implemented into a xylitol production system. Cell-recycle operation maintained concentration of the recombinant cells high inside a bioreactor. Final dry cell mass of 22.0 g/l, 116 g/l xylitol concentration, 2.34 g/l h overall xylitol productivity were obtained in cell-recycle fermentation supplemented with xylose and yeast extract solution, which were equivalent to 2.3-, 5.8- and 3.8-fold increases compared with the corresponding values of batch-type xylitol production parameters.     

Formation of xylitol in Candida guilliermondii 2581 culture

The yeast strain Candida guilliermondii 2581 was chosen for its ability to produce xylitol in media with high concentrations of xylose. The rate of xylitol production at a xylose concentration of 150 g/l is 1.25 g/l per h; the concentration of xylitol after three days of cultivation is 90 g/l; and the relative xylitol yield is 0.6 g per g substrate consumed. The growth conditions were found that resulted in the maximum relative xylitol yield with complete consumption of the sugar: xylose concentration, 150 g/l; pH 6.0; and shaking at 60 rpm. It was shown that the growth under conditions of limited aeration favors the reduction of xylose.     

Batch xylitol production by<Emphasis Type="Italic"> Candida guilliermondii</Emphasis> FTI 20037 from sugarcane bagasse hemicellulosic hydrolyzate at controlled pH values

Batch fermentation of sugarcane bagasse hemicellulosic hydrolyzate by the yeast Candida guilliermondii FTI 20037 was performed using controlled pH values (3.5, 5.5, 7.5). The maximum values of xylitol volumetric productivity (Q _p=0.76 g/l h) and xylose volumetric consumption (Q _s=1.19 g/l h) were attained at pH 5.5. At pH 3.5 and 7.5 the Q _p value decreased by 66 and 72%, respectively. Independently of the pH value, Y _x/s decreased with the increase in Y _p/s suggesting that the xylitol bioconversion improves when the cellular growth is limited. At the highest pH value (7.5), the maximum specific xylitol production value was the lowest (q _pmax=0.085 g/l h.), indicating that the xylose metabolism of the yeast was diverted from xylitol formation to cell growth.List of symbols P _max xylitol concentration (g/l) - Q _x volumetric cell production rate (g/l h) - Q _s volumetric xylose uptake rate (g/l h) - Q _p volumetric xylitol production rate (g/l h) - q _pmax specific xylitol production (g/g h) - q _smax specific xylose uptake rate (g/g h) - _max specific cell growth rate (h^–1) - Y _p/s xylitol yield coefficient, g xylitol per g xylose consumed (g/g) - Y _p/x xylitol yield coefficient, g xylitol per g dry cell mass produced (g/g) - Y _x/s cell yield coefficient, g dry cell mass per g xylose consumed (g/g) - _cell percentage of the cell yield from the theoretical value (%) - _xylitol percentage of xylitol yield from the theoretical value (%)     

Xylitol Production by a Culture ofCandida guilliermondii2581

The yeast strain Candida guilliermondii2581 was chosen for its ability to produce xylitol in media with high concentrations of xylose. The rate of xylitol production at a xylose concentration of 150 g/l is 1.25 g/l per h; the concentration of xylitol after three days of cultivation is 90 g/l; and the relative xylitol yield is 0.6 g per g substrate consumed. The growth conditions were found that resulted in the maximum relative xylitol yield with complete consumption of the sugar: xylose concentration, 150 g/l; pH 6.0; and shaking at 60 rpm. It was shown that the growth under conditions of limited aeration favors the reduction of xylose.     

Xylitol production by <Emphasis Type="Italic">Candida tropicalis</Emphasis> in a chemically defined medium

A chemically defined medium that included urea (5 g l(-1)) as a nitrogen source and various vitamins was substituted for a complex medium containing yeast extract (10 g l(-1)) in the production of xylitol by Candida tropicalis. In a fed-batch culture with the chemically defined medium, 237 g xylitol l(-1) was produced from 270 g xylose l(-1) after 120 h. The volumetric rate of xylitol production and the xylitol yield from xylose were 2 g l(-1) h(-1) and 89%, respectively. These values were about 5% lower and 4% higher, respectively, than those obtained using the complex medium. These results indicate that xylitol can be produced effectively in a chemically defined medium.     

Fermentation behavior of an osmotolerant yeast D. hansenii for Xylitol production

Realizing the importance of xylitol as a high‐valued compound that serves as a sugar substitute, a new, one step thin layer chromatographic procedure for quick, reliable, and efficient determination of xylose and xylitol from their mixture was developed. Two hundred and twenty microorganisms from the laboratory stock cultures were screened for their ability to produce xylitol from D ‐xylose. Amongst these, an indigenous yeast isolate no.139 (SM‐139) was selected and identified as Debaryomyces hansenii on the basis of morphological and biochemical characteristics and (26S) D1/D2 r DNA region sequencing. Debaryomyces hansenii produced 9.33 gL^?1 of xylitol in presence of 50.0 gL^?1 of xylose in 84 h at pH 5.5, 30°C, 200 rpm. In order to utilize even higher concentrations of xylose for maximum xylitol production, a xylose enrichment technique was developed. The strain of Debaryomyces hansenii was obtained through xylose enrichment technique in a statistically optimized medium containing 0.3% yeast extract, 0.2% peptone, 0.03% MgSO₄.7H₂O along with 1% methanol. The culture was inoculated with 6% inoculum and incubated at 30°C and 250 rpm. A yield of 0.6 gg^?1 was obtained with a xylitol volumetric productivity of 0.65 g/L h^?1 in the presence of 200 gL^?1 of xylose although up to 300 gL^?1 of xylose could be tolerated through batch fermentation. Through this technique, even higher concentrations of xylose as substrate could be potentially utilized for maximum xylitol production. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

Effects of environmental conditions on production of xylitol byCandida boidinii

Candida boidinii NRRL Y-17213 produced more xylitol thanC. magnolia (NRRL Y-4226 and NRRL Y-7621),Debaryomyces hansenii (C-98 M-21, C-56 M-9 and NRRL Y-7425), orPichia (Hansenula) anomala (NRRL Y-366). WithC. boidinii, highest xylitol productivity was at pH 7 but highest yield was at pH 8, using 5 g urea and 5 g Casamino acids/I. Decreasing the aeration rate decreased xylose consumption and cell growth but increased the xylitol yield. When an initial cell density of 5.1 g/l was used instead of 1.3 g/l, xylitol yield and the specific xylitol production rate doubled. Substrate concentration had the greatest effect on xylitol production; increasing xylose concentration 7.5-fold (to 150 g/l) gave a 71-fold increase in xylitol production (53 g/l) and a 10-fold increase in xylitol/ethanol ratio. The highest xylitol yield (0.47 g/g), corresponding to 52% of the theoretical yield, was obtained with 150 g xylose/l after 14 days. Xylose at 200 g/l inhibited xylitol production.E. Vandeska and S. Kuzmanova were and S. Amartey and T. Jeffries are with the Forest Products Laboratory, Institute for Microbial and Biochemical Technology, 1 Gifford Pinchot Drive, Madison, WI 53703, USA. E. Vandeska and S. Kuzmanova are now with the Faculty of Technology and Metallurgy, Rudjer Boskovic 16, 91000 Skopje, Macedonia     

Increase of xylitol yield by feeding xylose and glucose in Candida tropicalis

Candida tropicalis, a strain isolated from the sludge of a factory manufacturing xylose, produced a high xylitol concentration of 131 g/l from 150 g/l xylose at 45 h in a flask. Above 150 g/l xylose, however, volumetric xylitol production rates decreased because of a lag period in cell growth. In fed-batch culture, the volumetric production rate and xylitol yield from xylose varied substantially with the controlled xylose concentration and were maximum at a controlled xylose concentration of 60 g/l. To increase the xylitol yield from xylose, feeding experiments using different ratios of xylose and glucose were carried out in a fermentor. The maximum xylitol yield from 300 g/l xylose was 91% at a glucose/xylose feeding ratio of 15%, while the maximum volumetric production rate of xylitol was 3.98 g l⁻¹ h⁻¹ at a glucose/xylose feeding ratio of 20%. Xylitol production was found to decrease markedly as its concentration rose above 250 g/l. In order to accumulate xylitol to 250 g/l, 270 g/l xylose was added in total, at a glucose/xylose feeding ratio of 15%. Under these conditions, a final xylitol production of 251 g/l, which corresponded to a yield of 93%, was obtained from 270 g/l xylose in 55 h. Received: 20 April 1998 / Received revision: 29 May 1998 / Accepted: 19 June 1998     

Fed-batch xylitol production with recombinantXYL-1-expressingSaccharomyees cerevisiae using ethanol as a co-substrate

The bioconversion of xylose into xylitol in fed-batch fermentation with a recombinantSaccharomyces cerevisiae strain, transformed with the xylose-reductase gene ofPichia stipitis, was studied. When only xylose was fed into the fermentor, the production of xylitol continued until the ethanol that had been produced during an initial growth phase on glucose, was depleted. It was concluded that ethanol acted as a redox-balance-retaining co-substrate. The conversion of high amounts of xylose into xylitol required the addition of ethanol to the feed solution. Under O₂-limited conditions, acetic acid accumulated in the fermentation broth, causing poisoning of the yeast at low extracellular pH. Acetic acid toxicity could be avoided by either increasing the pH from 4.5 to 6.5 or by more effective aeration, leading to the further metabolism of acetic acid into cell mass. The best xylitol/ethanol yield, 2.4 gg^–1 was achieved under O₂-limited conditions. Under anaerobic conditions ethanol could not be used as a co-substrate, because the cell cannot produce ATP for maintenance requirements from ethanol anaerobically. The specific rate of xylitol production decreased with increasing aeration. The initial volumetric productivity increased when xylose was added in portions rather than by continuous feeding, due to a more complete saturation of the transport system and the xylose reductase enzyme.     

Variables that affect xylitol production from sugarcane bagasse hydrolysate in a zeolite fluidized bed reactor

The operational conditions for xylitol production by fermentation of sugarcane bagasse hydrolysate in a fluidized bed reactor with cells immobilized on zeolite were evaluated. Fermentations were carried out under different conditions of air flowrate (0.0125-0.0375 vvm), zeolite mass (100-200 g), initial pH (4-6), and xylose concentration (40-60 g/L), according to a 2(4) full factorial design. The air flowrate increase resulted in a metabolic deviation from product to biomass formation. On the other hand, the pH increase favored both the xylitol yield (Y(P/S)) and volumetric productivity (Q(P)), and the xylose concentration increase positively influenced the xylitol concentration. The best operational conditions evaluated were based on the use of an air flowrate of 0.0125 vvm, 100 g of zeolite, pH 6, and xylose concentration of 60 g/L. Under these conditions, 38.5 g/L of xylitol were obtained, with a Y(P/S) of 0.72 g/g, Q(P) of 0.32 g/L.h, and cell retention of 25.9%.     

The influence of hexoses addition on the fermentation of d-xylose in Debaryomyces hansenii under continuous cultivation

The effect of hexoses (glucose and galactose) addition to the feed xylose mineral medium of Debaryomyces hansenii chemostat cultures grown at a constant dilution rate of 0.055 h⁻¹ was studied. Xylitol was the major product detected amongst all tested conditions. The maximal values for xylitol yield and volumetric productivity (0.56 gg⁻¹ xylose and 0.21 gl⁻¹h⁻¹, respectively) were obtained for a glucose/xylose feeding ratio of 10%, showing that the addition of small amounts of glucose, but not galactose, enhanced the xylitol production. A xylitol yield increase of 30%, compared with the sole xylose-containing feed medium, was observed. It was found that the oxygen requirement for D. hansenii growth is lower under glucose compared with xylose. Ethanol and glycerol were only produced for glucose/xylose feeding ratio above 30%. The byproducts accumulation was correlated with glucose metabolism, because a direct relationship between the increase of ethanol (and glycerol) concentration and the increase of glucose in the feed medium was found.     

Effects of NADH-preferring xylose reductase expression on ethanol production from xylose in xylose-metabolizing recombinant Saccharomyces cerevisiae

Efficient conversion of xylose to ethanol is an essential factor for commercialization of lignocellulosic ethanol. To minimize production of xylitol, a major by-product in xylose metabolism and concomitantly improve ethanol production, Saccharomyces cerevisiae D452-2 was engineered to overexpress NADH-preferable xylose reductase mutant (XR(MUT)) and NAD?-dependent xylitol dehydrogenase (XDH) from Pichia stipitis and endogenous xylulokinase (XK). In vitro enzyme assay confirmed the functional expression of XR(MUT), XDH and XK in recombinant S. cerevisiae strains. The change of wild type XR to XR(MUT) along with XK overexpression led to reduction of xylitol accumulation in microaerobic culture. More modulation of the xylose metabolism including overexpression of XR(MUT) and transaldolase, and disruption of the chromosomal ALD6 gene encoding aldehyde dehydrogenase (SX6(MUT)) improved the performance of ethanol production from xylose remarkably. Finally, oxygen-limited fermentation of S. cerevisiae SX6(MUT) resulted in 0.64 g l?1 h?1 xylose consumption rate, 0.25 g l?1 h?1 ethanol productivity and 39% ethanol yield based on the xylose consumed, which were 1.8, 4.2 and 2.2 times higher than the corresponding values of recombinant S. cerevisiae expressing XR(MUT), XDH and XK only.     

Influence of cultivation conditions on xylose-to-xylitol bioconversion by a new isolate of Debaryomyces hansenii

About 270 yeast isolates were screened for xylitol production using xylose as the sole carbon source. The best isolate, Debaryomyces hansenii UFV-170, released 5.84 g L(-1) xylitol from 10 g L(-1) xylose after 24 h, corresponding to a yield of xylitol on consumed substrate (Y(P/S)) of 0.54 g g(-1). This strain was cultivated batch-wise at variable starting concentrations of xylose (S(o)) and biomass (X(o)) and agitation intensity, in order to improve xylitol production and to evaluate, through simple carbon balances, the influence of these conditions on xylose metabolism. Under the best microaerobic conditions (S(o) = 53 g L(-1), X(o) = 1.4 g L(-1), 200 rpm), xylitol production reached 37.0 g L(-1), corresponding to xylitol volumetric productivity of 1.0 g L(-1)h(-1), specific productivity of 0.22 g g(-1)h(-1) and Y(P/S) = 0.76 g g(-1). Almost 83% of xylose was consumed for xylitol production, the rest being consumed for growth, while respiration was negligible. The new isolate appeared to be a promising alternative for industrial xylitol bioproduction.     

Xylitol production from sugarcane bagasse hydrolyzate in fluidized bed reactor. Effect of air flowrate

Cells of Candida guilliermondii immobilized onto porous glass spheres were cultured batchwise in a fluidized bed bioreactor for xylitol production from sugarcane bagasse hemicellulose hydrolyzate. An aeration rate of only 25 mL/min ensured minimum yields of xylose consumption (0.60) and biomass production (0.14 g(DM)/g(Xyl)), as well as maximum xylitol yield (0.54 g(Xyt)/g(Xyl)) and ratio of immobilized to total cells (0.83). These results suggest that cell metabolism, although slow because of oxygen limitation, was mainly addressed to xylitol production. A progressive increase in the aeration rate up to 140 mL/min accelerated both xylose consumption (from 0.36 to 0.78 g(Xyl)/L.h) and xylitol formation (from 0.19 to 0.28 g(Xyt)/L.h) but caused the fraction of immobilized to total cells and the xylitol yield to decrease up to 0.22 and 0.36 g(Xyt)/g(Xyl), respectively. The highest xylitol concentration (17.0 g(Xyt)/L) was obtained at 70 mL/min, but the specific xylitol productivity and the xylitol yield were 43% and 22% lower than the corresponding values obtained at the lowest air flowrate, respectively. The concentrations of consumed substrates and formed products were used in material balances to evaluate the xylose fractions consumed by C. guilliermondii for xylitol production, complete oxidation through the hexose monophosphate shunt, and cell growth. The experimental data collected at variable oxygen level allowed estimating a P/O ratio of 1.35 mol(ATP)/mol(O) and overall ATP requirements for biomass growth and maintenance of 3.4 mol(ATP)/C-mol(DM).     

Reduced oxidative pentose phosphate pathway flux in recombinant xylose-utilizing Saccharomyces cerevisiae strains improves the ethanol yield from xylose

In recombinant, xylose-fermenting Saccharomyces cerevisiae, about 30% of the consumed xylose is converted to xylitol. Xylitol production results from a cofactor imbalance, since xylose reductase uses both NADPH and NADH, while xylitol dehydrogenase uses only NAD(+). In this study we increased the ethanol yield and decreased the xylitol yield by lowering the flux through the NADPH-producing pentose phosphate pathway. The pentose phosphate pathway was blocked either by disruption of the GND1 gene, one of the isogenes of 6-phosphogluconate dehydrogenase, or by disruption of the ZWF1 gene, which encodes glucose 6-phosphate dehydrogenase. Decreasing the phosphoglucose isomerase activity by 90% also lowered the pentose phosphate pathway flux. These modifications all resulted in lower xylitol yield and higher ethanol yield than in the control strains. TMB3255, carrying a disruption of ZWF1, gave the highest ethanol yield (0.41 g g(-1)) and the lowest xylitol yield (0.05 g g(-1)) reported for a xylose-fermenting recombinant S. cerevisiae strain, but also an 84% lower xylose consumption rate. The low xylose fermentation rate is probably due to limited NADPH-mediated xylose reduction. Metabolic flux modeling of TMB3255 confirmed that the NADPH-producing pentose phosphate pathway was blocked and that xylose reduction was mediated only by NADH, leading to a lower rate of xylose consumption. These results indicate that xylitol production is strongly connected to the flux through the oxidative part of the pentose phosphate pathway.     

酵母发酵蔗渣半纤维素水解物生产木糖酶

采用二次正交旋转组合设计研究了蔗渣半纤维素水解过程中硫酸浓度与液 固比对木糖收率的影响。回归分析表明 ,这两个因素与木糖的收率之间存在显著的回归关系。通过回归方程优化水解条件 ,当硫酸浓度 2 .4g L ,液 固 =6 .2 ,在蒸汽压力 2 .5× 10 4Pa的条件下水解 2 .5h ,10 0g蔗渣可水解生成木糖约 2 4g。大孔树脂吸附层析处理蔗渣半纤维素水解物 ,能有效地减少其中的酵母生长抑制物含量 ,显著改善水解物的发酵性能。用大孔树脂在pH 2条件下处理过的蔗渣半纤维素水解物作基质 ,含木糖 2 0 0g L ,产木糖醇酵母菌株CandidatropicalisAS2 .1776发酵 110h耗完基质中的木糖 ,生成木糖醇 12 7g L ,产物转化率 0 .6 4(木糖醇g 木糖g) ,产物生成速率 1.15g L·h .