Improving the Mould and Blue-Stain-Resistance of Bamboo through Acidic Hydrolysis

Bamboo is much more easily attacked by fungus compared with wood, resulting in shorter service life and higher loss in storage and transportation. It has been long accepted that the high content of starch and sugars in bamboo is mainly responsible for its low mould resistance. In this paper, acetic acid, propionic acid, oxalic acid, citric acid, and hydrochloric acid were adopted to hydrothermally hydrolyze the starch in bamboo, with the aims to investigate their respective effect on the mould and blue-stain resistance of bamboo, and the optimized citric acid in different concentrations were studied. The starch content, glucose yields, weight loss, and colour changes of solid bamboo caused by the different acidic hydrolysis were also compared. The results indicated that weak acidic hydrolysis treatment was capable of improving mould-resistant of bamboo. The mould resistance increased with the increased concentration of citric acid. Bamboo treated with citric acid in the concentration of 10% could reduce the infected area ranging to 10–17%, the growth rating of which could reach 1 resistance. The content of soluble sugar and starch remained in bamboo decreased significantly from 43 mg/g to 31 mg/g and 46 mg/g to 23 mg/g, respectively, when the citric acid concentration varied from 4% to 10%. Citric acid treatments of 10% also caused a greatest surface colour change and weight loss. The results in this study demonstrated citric acid treatment can effectively reduce the starch grain and soluble sugars content and improve mould resistance of bamboo, which can be attributed to the reduction of starch grain and soluble carbohydrates (such as glucose, fructose, and sucrose, etc.) in bamboo.

Transformations of 3,7-interphenylene 11-deoxyprostanoid formyl precursors in the acidic medium

It has been shown that the formyl precursors of 3,7-interphenylene 11-deoxyprostanoids, formed during acidic hydrolysis of the corresponding acetals, can undergo isomerization (disproportionation) in the acidic medium to give 2-(arylalkyl)-3-(hydroxymethyl)cyclopent-2-ene-1-ones – the synthons for prostanoids and phytoprostanes of the series B. Acetal precursors of 3,7-interphenylene 11-deoxyprostaglandin analogues with electron-donating alkoxy substituent in position 3′ of the aromatic fragment in the α-chain under similar conditions hydrolyze with the formation of formyl derivatives that spontaneously cyclize to produce 2,3,4,9-tetrahydro-1H-cyclopenta[b]naphthalene-1-ones.

Bioconversion of Industrial Cassava Solid Waste (Onggok) to Bioethanol Using a Saccharification and Fermentation process

Cassava solid waste (Onggok) is a by-product of the starch industry containing a lot of fiber, especially cellulose and hemicellulose. It has the potential to be converted to bioethanol. This work aimed to evaluate the effect of feedstocks ratio for the optimal bioethanol production via enzymatic and acidic hydrolysis process in a batch fermentation process. The effect of alpha-amylase and glucoamylase activities was studied. The sulfuric acid concentrations in the hydrolysis process in converting cassava into reducing sugar were also investigated. The reducing sugar was then fermented to produce ethanol. Enzymatic and chemical hydrolysis was carried out with the ratio of onggok(g)/water(L), 50/1, 75/1, and 100/1 (w/v). In the enzymatic hydrolysis, 22.5, 45, and 67.5 KNU (Kilo Novo alpha-amylase Unit) for liquefaction; and 65, 130, and 195 GAU (Glucoamylase Unit) for saccharification, respectively of enzymes were applied. The liquefaction was carried out at 90-100⁰C for 2 hours. The saccharification was executed at 65 ⁰C for 4 hours. Meanwhile, the acidic hydrolysis operating condition was at 90-100 ⁰C for 3 hours. The fermentation was performed at pH 4.5 for 3 days. Fourier Transform Infra-Red (FTIR) analysis was conducted to evaluate the hydrolysis process. The highest ethanol was yielded in the fermentation at 8.89% with the ratio of onggok to water 100:1, 67.5 KNU of alpha-amylase, and 195 GAU of glucoamylase. Ethanol was further purified utilizing fractional distillation. The final ethanol concentration was at 93-94%.

Mass Spectrometric Quantification of the Antimicrobial Peptide Pep19-2.5 with Stable Isotope Labeling and Acidic Hydrolysis

Sepsis is the number one cause of death in intensive care units. This life-threatening condition is caused by bacterial infections and triggered by endotoxins of Gram-negative bacteria that leads to an overreaction of the immune system. The synthetic anti-lipopolysaccharide peptide Pep19-2.5 is a promising candidate for the treatment of sepsis as it binds sepsis-inducing lipopolysaccharides and thus prevents initiation of septic shock. For clinical evaluation precise quantification of the peptide in blood and tissue is required. As the peptide is not extractable from biological samples by commonly used methods there is a need for a new analysis method that does not rely on extraction of the peptide. In order to quantify the peptide by mass spectrometry, the peptide was synthesized containing 13C9,15N1-labeled phenylalanine residues. This modification offers high stability during acidic hydrolysis. Following acidic hydrolysis of the samples, the concentration of 13C9,15N1-labeled phenylalanine determined by LC-MS could be unambiguously correlated to the content of Pep19-2.5. Further experiments validated the accuracy of the data. Moreover, the quantification of Pep19-2.5 in different tissues (as studied in Wistar rats) was shown to provide comparable results to the results obtained with radioactively-labeled (14C) Pep19-2.5- Radioactive labeling is considered as the gold standard for quantification of compounds that refrain from reliable extraction methods. This novel method represents a valuable procedure for the determination of Pep19-2.5 and sticky peptides with unpredictable extraction properties in general.

Spanish Poplar Biomass as a Precursor for Nanocellulose Extraction

The effect of acidic hydrolysis duration on nanocellulose size, morphology, and proper ties was investigated, which opens up a whole new horizon of versatility in poplar applications. This study aimed to examine Spanish poplar wastes as raw material to extract crystalline nanocellulose (CNC), which substantiates the importance of poplar wastes. Wastes were pulped using 1 L of 10% NaOH (wt./wt.) solution, and bleached several times by NaClO2; afterwards, white wastes were subjected to acidic hydrolysis by 60% H2SO4 for either 5, 10, or 15 min. Microcrystalline cellulose (MCC) underwent a similar hydrolysis protocol as poplar as control. TEM, IR, and XRD characterization techniques were performed. Poplar based nanocellulose sized 219 nm length and 69 nm width after 15 min acidic hydrolysis. MCC yielded 122 nm length and 12 nm width crystals after 10 min acidic hydrolysis. Hydrolysis resulted in a drastic change and intense peaks at 3500 and 2900 cm−1 for nanocellulose. Although pre-hydrolysis fiber treatment was not influencial on the crystallinity of poplar, acidic hydrolysis remarkably raised the crystallinity index (CI) by 7–8%. The more hydrolysis duration was prolonged, the size of the resulting crystal (whisker) decreased, and the aspect ratio increased. Hydrolysis was more impactful on MCC than poplar. However, for future work, it seems that longer duration of pulping and bleaching could have significantly removed unwanted components (hemicellulose and lignin), showcased in IR and XRD, and hence smoothened the following hydrolysis.

Optimised Fractionation of Brewer’s Spent Grain for a Biorefinery Producing Sugars, Oligosaccharides, and Bioethanol

Brewer’s spent grain (BSG) is the main by-product of the beer brewing process. It has a huge potential as a feedstock for bio-based manufacturing processes to produce high-value bio-products, biofuels, and platform chemicals. For the valorisation of BSG in a biorefinery process, efficient fractionation and bio-conversion processes are required. The aim of our study was to develop a novel fractionation of BSG for the production of arabinose, arabino-xylooligomers, xylose, and bioethanol. A fractionation process including two-step acidic and enzymatic hydrolysis steps was investigated and optimised by a response surface methodology and a desirability function approach to fractionate the carbohydrate content of BSG. In the first acidic hydrolysis, high arabinose yield (76%) was achieved under the optimised conditions (90 °C, 1.85 w/w% sulphuric acid, 19.5 min) and an arabinose- and arabino-xylooligomer-rich supernatant was obtained. In the second acidic hydrolysis, the remaining xylan was solubilised (90% xylose yield) resulting in a xylose-rich hydrolysate. The last, enzymatic hydrolysis step resulted in a glucose-rich supernatant (46 g/L) under optimised conditions (15 w/w% solids loading, 0.04 g/g enzyme dosage). The glucose-rich fraction was successfully used for bioethanol production (72% ethanol yield by commercial baker’s yeast). The developed and optimised process offers an efficient way for the value-added utilisation of BSG. Based on the validated models, the amounts of the produced sugars, the composition of the sugar streams and solubilised oligo-saccharides are predictable and variable by changing the reaction conditions of the process.

In-peptide amino acid racemization via inter-residue oxazoline intermediates during acidic hydrolysis

Isopedopeptins are antibiotic cyclic lipodepsipeptides containing the subsequence L-Thr—L-2,3-diaminopropanoic acid—D-Phe—L-Val/L-3-hydroxyvaline. Acidic hydrolysis of isopedopeptins in D2O showed the D-Phe residues to racemize extensively in peptides with L-3-hydroxyvaline but not in peptides with L-Val. Similarly, one Leu residue in pedopeptins, which are related peptides containing the subsequence Leu—2,3-diaminopropanoic acid—Leu—L-Val/L-3-hydroxyvaline, was found to racemize in peptides with L-3-hydroxyvaline. Model tetrapeptides, L-Ala—L-Phe—L-Val/3-hydroxyvaline—L-Ala, gave the corresponding results, i.e. racemization of L-Phe only when linked to a L-3-hydroxyvaline. We propose the racemization to proceed via an oxazoline intermediate involving Phe/Leu and the L-3-hydroxyvaline residues. The 3-hydroxyvaline residue may form a stable tertiary carbocation by loss of the sidechain hydroxyl group as water after protonation. Elimination of the Phe/Leu H-2 and ring-closure from the carbonyl oxygen onto the carbocation results in the suggested oxazoline intermediate. The reversed reaction leads to either retained or inversed configuration of Phe/Leu. Such racemization during acidic hydrolysis may occur whenever a 3-hydroxyvaline residue or any amino acid that can form a stable carbocation on the C-3, is present in a peptide. The proposed mechanism for racemization was supported by incorporation of 18O in the 3-hydroxyvaline sidechain when the acidic hydrolysis was performed in H2O/H218O (1:1). The 2,3-diaminopropanoic residues of isopedopeptins and pedopeptins were also found to racemize during acidic hydrolysis, as previously described. Based on the results, the configuration of the Leu and 2,3-diaminopropanoic acid residues of the pedopeptins were reassigned to be L-Leu and D-Leu, and 2 × L-2,3-diaminopropanoic acid.

Kinetic and Mechanistic Studies of Acidic Hydrolysis of Goniothalamin

The acidic hydrolysis of goniothalamin was studied on the spectrophotometric kinetic study at different concentration of hydrochloric acid and temperature to determine the stability of the compound. Stability tests were performed using UV-VIS detection. This is a two-step reaction that involves formation of intermediate product. Rate constant of reactant forming intermediate product obeyed pseudo-first-order kinetic, while the second step to form final product is independent on the concentration of HCl. The structure of final products was identified by NMR and MS. The acidic hydrolysis pathway was proposed to involve the opening of lactone ring, followed by dehydration and formation of a double bond.