Effects of acid hydrolysis waste liquid recycle on preparation of microcrystalline cellulose

Large amounts of acidic waste are produced on the industrial scale during hydrolysis of partially amorphous cellulose to produce microcrystalline cellulose (MCC). The essential disposal and treatment of this highly acidic liquid wastes the acid feedstock and increases the production cost. To maximize the use of acid without sacrificing the MCC product quality, this project reports a successful attempt to recycle the acid hydrolysis waste liquid, focusing on the impact of waste recycling on MCC morphology and reducing sugar in the hydrolysate. The results showed that when the waste liquid is recycled 1-5 times, no metal accumulation occurred while cellulose particles remained intact, maintaining their shape and size. Their extent of crystallinity remained nearly constant, even increasing slightly with up to three cycles. The concentration of reducing sugar showed growth when recycling the waste liquid up to three times, although not quite to the levels that would allow for its cost-effective fermentation. The acid amount to be added at the start of each cycle was near 50% of that used on the first stage

Liver Toxicity of Rare Ginsenosides in Rats after 13 Weeks of Oral Exposure

Objective: This study aims to evaluate the hepatotoxicity of rare ginsenosides in rats after 90 days of oral administration using heat-transformed rare ginsenosides as the primary material. Methods: A total of 48 male and female rats were randomly assigned into four groups: High-dose rare ginsenosides (600 mg/kg), medium-dose rare ginsenosides (200 mg/kg), low-dose rare ginsenosides (60 mg/kg), and a blank control group. After 90 days of oral gavage treatment, ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) was employed for metabolomic analysis of rat serum and flow cytometry analysis of liver apoptosis to evaluate the potential liver damage comprehensively in rats. Results: A significant difference in hepatocyte apoptotic rate was observed between the high-dose group and the control group in both male and female rats (P0.05). However, 23 differential metabolites, such as histidine, glutamate, proline and arginine were identified in the serum of female rats in the high-dose group, affecting the histidine and urea cycle metabolic pathways and causing hyperammonemia and liver damage. Ten differential metabolites affecting the alpha-linolenic acid and linoleic acid metabolic pathways were found in male rats, such as linoleic acid and arachidonic acid. High concentrations of arachidonic acid showed inflammatory and toxic effects, which could be absorbed into the portal vein system through blood and cause liver damage. Conclusion: High-dose rare ginsenosides mainly cause slight liver damage in male and female rats mainly due to the changes of histidine, α-linolenic acid and linoleic acid metabolic pathways. Hence, no adverse liver effects were observed at doses less than 200 mg/kg in both male and female rats

Expression of Codon-Optimized Plant Glycosyltransferase UGT72B14 in Escherichia coli Enhances Salidroside Production

Salidroside, a plant secondary metabolite in Rhodiola, has been demonstrated to have several adaptogenic properties as a medicinal herb. Due to the limitation of plant source, microbial production of salidroside by expression of plant uridine diphosphate glycosyltransferase (UGT) is promising. However, glycoside production usually remains hampered by poor expression of plant UGTs in microorganisms. Herein, we achieved salidroside production by expression of Rhodiola UGT72B14 in Escherichia coli (E. coli) and codon optimization was accordingly applied. UGT72B14 expression was optimized by changing 278 nucleotides and decreasing the G+C content to 51.05% without altering the amino acid sequence. The effect of codon optimization on UGT72B14 catalysis for salidroside production was assessed both in vitro and in vivo. In vitro, salidroside production by codon-optimized UGT72B14 is enhanced because of a significantly improved protein yield (increased by 4.8-fold) and an equivalently high activity as demonstrated by similar kinetic parameters (KM and Vmax), compared to that by wild-type protein. In vivo, both batch and fed-batch cultivation using the codon-optimized gene resulted in a significant increase in salidroside production, which was up to 6.7 mg/L increasing 3.2-fold over the wild-type UGT72B14

Effects of acid hydrolysis waste liquid recycle on preparation of microcrystalline cellulose

Large amounts of acidic waste are produced on the industrial scale during hydrolysis of partially amorphous cellulose to produce microcrystalline cellulose (MCC). The essential disposal and treatment of this highly acidic liquid wastes the acid feedstock and increases the production cost. To maximize the use of acid without sacrificing the MCC product quality, this project reports a successful attempt to recycle the acid hydrolysis waste liquid, focusing on the impact of waste recycling on MCC morphology and reducing sugar in the hydrolysate. The results showed that when the waste liquid is recycled 1-5 times, no metal accumulation occurred while cellulose particles remained intact, maintaining their shape and size. Their extent of crystallinity remained nearly constant, even increasing slightly with up to three cycles. The concentration of reducing sugar showed growth when recycling the waste liquid up to three times, although not quite to the levels that would allow for its cost-effective fermentation. The acid amount to be added at the start of each cycle was near 50% of that used on the first stage

Image_1_A high trans-zeatin nucleoside concentration in corms may promote the multileaf growth of Amorphophallus muelleri.tif

Amorphophallus muelleri has a multileaf growth pattern different from that of other konjacs; however, the hormonal mechanism underlying this phenomenon is not clear. In this study, the levels of hormones closely related to the sprouting of the axillary bud, including five types of cytokinins, indole-3-acetic acid (IAA) and abscisic acid (ABA) were measured. In the second leaf sprouting stage, the content of trans-zeatin riboside (tZR) in corms increased more than 5000-fold over that in the dormancy period. Surprisingly, although the expression of CYP735A1 and CYP735A2, which synthesize the precursors for tZR was elevated at the second leaf sprouting stage, the expression of IPTs, which have key roles in cytokinin biosynthesis, did not change significantly. In addition, most cytokinin contents in leaves during the same period were significantly lower than those in corms. We speculate that the high cytokinin contents in the corms may not biosynthesized de novo in corms. In addition, the IAA content in the corms also considerably increased during the second leaf sprouting stage. Indole-3-acetaldehyde oxidase (AO1) and auxin efflux carrier PIN1A, presented relatively high expression levels in the same period. In contrast, ABA content, and the expression of NCED1, a rate-limiting enzyme in ABA biosynthesis, were suppressed at the second leaf sprouting stage. It is worth mentioning that N6-(Δ2-isopentenyl) adenosine (iP)-type cytokinins have a high content in corms in the dormant period that significantly decreases after the first leaf sprouting stage, which is completely different from the trend of tZR. By treating dormant corms with iP, the percentage of multibud plants increased, and the growth performance in terms of bud and root length was significantly higher than those of the control. This implies that iP-type cytokinins tend to play a role in promoting first seedling sprouting. Furthermore, there was a remarkable increase of the IAA content in both corms and roots under iP treatment but an inhibitory effect in buds. We speculate that the increase in the IAA content induced by iP is tissue specific. These results will assist in the understanding of the role of hormones, especially cytokinins, in the multileaf growth type of konjac.</p

Table_1_A high trans-zeatin nucleoside concentration in corms may promote the multileaf growth of Amorphophallus muelleri.xlsx

Amorphophallus muelleri has a multileaf growth pattern different from that of other konjacs; however, the hormonal mechanism underlying this phenomenon is not clear. In this study, the levels of hormones closely related to the sprouting of the axillary bud, including five types of cytokinins, indole-3-acetic acid (IAA) and abscisic acid (ABA) were measured. In the second leaf sprouting stage, the content of trans-zeatin riboside (tZR) in corms increased more than 5000-fold over that in the dormancy period. Surprisingly, although the expression of CYP735A1 and CYP735A2, which synthesize the precursors for tZR was elevated at the second leaf sprouting stage, the expression of IPTs, which have key roles in cytokinin biosynthesis, did not change significantly. In addition, most cytokinin contents in leaves during the same period were significantly lower than those in corms. We speculate that the high cytokinin contents in the corms may not biosynthesized de novo in corms. In addition, the IAA content in the corms also considerably increased during the second leaf sprouting stage. Indole-3-acetaldehyde oxidase (AO1) and auxin efflux carrier PIN1A, presented relatively high expression levels in the same period. In contrast, ABA content, and the expression of NCED1, a rate-limiting enzyme in ABA biosynthesis, were suppressed at the second leaf sprouting stage. It is worth mentioning that N6-(Δ2-isopentenyl) adenosine (iP)-type cytokinins have a high content in corms in the dormant period that significantly decreases after the first leaf sprouting stage, which is completely different from the trend of tZR. By treating dormant corms with iP, the percentage of multibud plants increased, and the growth performance in terms of bud and root length was significantly higher than those of the control. This implies that iP-type cytokinins tend to play a role in promoting first seedling sprouting. Furthermore, there was a remarkable increase of the IAA content in both corms and roots under iP treatment but an inhibitory effect in buds. We speculate that the increase in the IAA content induced by iP is tissue specific. These results will assist in the understanding of the role of hormones, especially cytokinins, in the multileaf growth type of konjac.</p

Image_2_A high trans-zeatin nucleoside concentration in corms may promote the multileaf growth of Amorphophallus muelleri.tif

Amorphophallus muelleri has a multileaf growth pattern different from that of other konjacs; however, the hormonal mechanism underlying this phenomenon is not clear. In this study, the levels of hormones closely related to the sprouting of the axillary bud, including five types of cytokinins, indole-3-acetic acid (IAA) and abscisic acid (ABA) were measured. In the second leaf sprouting stage, the content of trans-zeatin riboside (tZR) in corms increased more than 5000-fold over that in the dormancy period. Surprisingly, although the expression of CYP735A1 and CYP735A2, which synthesize the precursors for tZR was elevated at the second leaf sprouting stage, the expression of IPTs, which have key roles in cytokinin biosynthesis, did not change significantly. In addition, most cytokinin contents in leaves during the same period were significantly lower than those in corms. We speculate that the high cytokinin contents in the corms may not biosynthesized de novo in corms. In addition, the IAA content in the corms also considerably increased during the second leaf sprouting stage. Indole-3-acetaldehyde oxidase (AO1) and auxin efflux carrier PIN1A, presented relatively high expression levels in the same period. In contrast, ABA content, and the expression of NCED1, a rate-limiting enzyme in ABA biosynthesis, were suppressed at the second leaf sprouting stage. It is worth mentioning that N6-(Δ2-isopentenyl) adenosine (iP)-type cytokinins have a high content in corms in the dormant period that significantly decreases after the first leaf sprouting stage, which is completely different from the trend of tZR. By treating dormant corms with iP, the percentage of multibud plants increased, and the growth performance in terms of bud and root length was significantly higher than those of the control. This implies that iP-type cytokinins tend to play a role in promoting first seedling sprouting. Furthermore, there was a remarkable increase of the IAA content in both corms and roots under iP treatment but an inhibitory effect in buds. We speculate that the increase in the IAA content induced by iP is tissue specific. These results will assist in the understanding of the role of hormones, especially cytokinins, in the multileaf growth type of konjac.</p

Image_3_A high trans-zeatin nucleoside concentration in corms may promote the multileaf growth of Amorphophallus muelleri.tif

Amorphophallus muelleri has a multileaf growth pattern different from that of other konjacs; however, the hormonal mechanism underlying this phenomenon is not clear. In this study, the levels of hormones closely related to the sprouting of the axillary bud, including five types of cytokinins, indole-3-acetic acid (IAA) and abscisic acid (ABA) were measured. In the second leaf sprouting stage, the content of trans-zeatin riboside (tZR) in corms increased more than 5000-fold over that in the dormancy period. Surprisingly, although the expression of CYP735A1 and CYP735A2, which synthesize the precursors for tZR was elevated at the second leaf sprouting stage, the expression of IPTs, which have key roles in cytokinin biosynthesis, did not change significantly. In addition, most cytokinin contents in leaves during the same period were significantly lower than those in corms. We speculate that the high cytokinin contents in the corms may not biosynthesized de novo in corms. In addition, the IAA content in the corms also considerably increased during the second leaf sprouting stage. Indole-3-acetaldehyde oxidase (AO1) and auxin efflux carrier PIN1A, presented relatively high expression levels in the same period. In contrast, ABA content, and the expression of NCED1, a rate-limiting enzyme in ABA biosynthesis, were suppressed at the second leaf sprouting stage. It is worth mentioning that N6-(Δ2-isopentenyl) adenosine (iP)-type cytokinins have a high content in corms in the dormant period that significantly decreases after the first leaf sprouting stage, which is completely different from the trend of tZR. By treating dormant corms with iP, the percentage of multibud plants increased, and the growth performance in terms of bud and root length was significantly higher than those of the control. This implies that iP-type cytokinins tend to play a role in promoting first seedling sprouting. Furthermore, there was a remarkable increase of the IAA content in both corms and roots under iP treatment but an inhibitory effect in buds. We speculate that the increase in the IAA content induced by iP is tissue specific. These results will assist in the understanding of the role of hormones, especially cytokinins, in the multileaf growth type of konjac.</p