Acrylamide and 5-Hydroxymethylfurfural in Synthetic Sugar Cane Syrup: Mitigation by Additives.

The ability of additives to reduce the formation of acrylamide in simulated sugar cane syrups was investigated. Organic acids, B vitamins, and inorganic salts were added individually and in combination to simulated thickened cane juice, and the mixtures were heated at 120 °C for 30 min. Calcium chloride (1%), citric acid (0.1%), and vitamin B3 (0.1%) were the most effective individual additives from each chemical family. The effects of CaCl2 (0-1%), citric acid (0-0.125%), and vitamin B3 (0-0.1125%), when added in combination, on the concentrations of acrylamide and hydroxymethylfurfural (HMF) were studied using a Box-Behnken design. Combinations of all three additives lowered the acrylamide production, but only the combination of citric acid and vitamin B3 had a significant synergistic effect. However, all these additives stimulated the production of HMF, and no significant interactive effect between pairs of additives on HMF production was observed. Calcium chloride stimulated the formation of HMF most strongly. These results indicate that certain combinations of these additives effectively reduce acrylamide formation, but they also lead to an increase in the formation of HMF in sugar syrup.

Acrylamide in non-centrifugal sugars and syrups.

BACKGROUND: Acrylamide in foods has been widely studied because of its possible carcinogenicity. Most of the foods investigated were prepared using low moisture and high temperature conditions. Non-centrifugal sugars (NCSs), which have been promoted as 'non-chemical' natural sweeteners, contain precursors of acrylamide and their production processes involved prolonged heating. The acrylamide content in 32 commercial NCSs from coconut, cane and palmyra palm purchased in Asian countries was investigated. Additionally, syrups (80 o Brix) produced from coconut and palmyra raw saps and cane juice were prepared by evaporation with prolonged heating (2.5 h to reach 100 °C, 1 h to increase to 110 °C, held at 110 °C for 30 min). The compositions and contents of sugars, amino acids and minerals, as well as the physical characteristics of the raw saps, juice and syrups, were determined. RESULTS: The acrylamide content of these 32 products ranged from < 15 to 4011 µg kg-1 . The raw saps and juice were mildly acidic (pH 5.14-5.66) and similar values were observed for their syrups (4.73-5.73). The contents of sucrose, fructose and glucose in the saps and juice from these plants were similar, whereas their compositions varied with respect to amino acids. The variation of the ornithine content was significant, demonstrating a striking influence on the extent of acrylamide formation (867-1564 µg kg-1 ) in the syrups prepared from these materials. CONCLUSION: The present study emphasizes the importance of a careful monitoring and control of the critical steps invloved in the manufacturing process of NCSs (particularly the evaporation phase), aiming to protect the health and safety of consumers. © 2021 Society of Chemical Industry.

Ethanol Production Potential of Ethanol-Tolerant Saccharomyces and Non-Saccharomyces Yeasts.

Four ethanologenic ethanol-tolerant yeast strains, Saccharomyces cerevisiae (ATKU132), Saccharomycodes ludwigii (ATKU47), and Issatchenkia orientalis (ATKU5-60 and ATKU5-70), were isolated by an enrichment technique in yeast extract peptone dextrose (YPD) medium supplemented with 10% (v/v) ethanol at 30°C. Among non-Saccharomyces yeasts, Sd. ludwigii ATKU47 exhibited the highest ethanol-tolerance and ethanol production, which was similar to S. cerevisiae ATKU132. The maximum range of ethanol concentrations produced at 37°C by S. cerevisiae ATKU132 and Sd. ludwigii ATKU47 from an initial D-glucose concentration of 20% (w/v) and 28% (w/v) sugarcane molasses were 9.46-9.82% (w/v) and 8.07-8.32% (w/v), respectively.

Discovery of potential protein biomarkers associated with sugarcane white leaf disease susceptibility using a comparative proteomic approach.

Sugarcane white leaf disease (SCWLD) is caused by phytoplasma, a serious sugarcane phytoplasma pathogen, which causes significant decreases in crop yield and sugar quality. The identification of proteins involved in the defense mechanism against SCWLD phytoplasma may help towards the development of varieties resistant to SCWLD. We investigated the proteomes of four sugarcane varieties with different levels of susceptibility to SCWLD phytoplasma infection, namely K88-92 and K95-84 (high), KK3 (moderate), and UT1 (low) by quantitative label-free nano-liquid chromatography-tandem mass spectrometry (nano LC-MS/MS). A total of 248 proteins were identified and compared among the four sugarcane varieties. Two potential candidate protein biomarkers for reduced susceptibility to SCWLD phytoplasma were identified as proteins detected only in UT1. The functions of these proteins are associated with protein folding, metal ion binding, and oxidoreductase. The candidate biomarkers could be useful for further study of the sugarcane defense mechanism against SCWLD phytoplasma, and in molecular and conventional breeding strategies for variety improvement.

A draft chromosome-scale genome assembly of a commercial sugarcane.

Sugarcane accounts for a large portion of the worlds sugar production. Modern commercial cultivars are complex hybrids of S. officinarum, S. spontaneum, and several other Saccharum species, resulting in an auto-allopolyploid with 8-12 copies of each chromosome. The current genome assembly gold standard is to generate a long read assembly followed by chromatin conformation capture sequencing to scaffold. We used the PacBio RSII and chromatin conformation capture sequencing to sequence and assemble the genome of a South East Asian commercial sugarcane cultivar, known as Khon Kaen 3. The Khon Kaen 3 genome assembled into 104,477 contigs totalling 7 Gb, which scaffolded into 56 pseudochromosomes containing 5.2 Gb of sequence. Genome annotation produced 242,406 genes from 30,927 orthogroups. Aligning the Khon Kaen 3 genome sequence to S. officinarum and S. spontaneum revealed a high level of apparent recombination, indicating a chimeric assembly. This assembly error is explained by high nucleotide identity between S. officinarum and S. spontaneum, where 91.8% of S. spontaneum aligns to S. officinarum at 94% identity. Thus, the subgenomes of commercial sugarcane are so similar that using short reads to correct long PacBio reads produced chimeric long reads. Future attempts to sequence sugarcane must take this information into account.

Pyrodextrins from waxy and normal tapioca starches: Molecular structure and in vitro digestibility.

Pyrodextrins were prepared from acidified waxy and normal tapioca starches (pH∼3) at 3 temperatures (130, 150 and 170 °C) and 3 times (1, 2 and 4 h) to determine their in vitro digestibility and molecular structure. Pyrodextrin from waxy tapioca starch produced at 170 °C/4 h had 5% higher total indigestible carbohydrate than pyrodextrin from normal tapioca starch (45.2 % and 40.4 %, respectively) as determined by a modified AOAC Method 2011.25. The low-molecular weight indigestible carbohydrate content at this condition was also higher for waxy tapioca starch than normal tapioca starch (40.6 % and 34.9 %, respectively). Gel permeation chromatography and nuclear magnetic resonance spectroscopy were used to study changes in molecular structure and correlate with digestibility of the pyrodextrins. Molecular size distribution indicated that waxy tapioca starch underwent thermal modification more readily than normal tapioca starch. Non α-1,4/α-1,6 glucosidic linkages were increased in the pyrodextrins with increasing in indigestible carbohydrate content.

The role of salicylic acid and benzothiadiazole in decreasing phytoplasma titer of sugarcane white leaf disease.

The objective of this research was to study the effect of Benzothiadiazole (BTH) and Salicylic acid (SA) on the systemic acquired resistance (SAR) of sugarcane the phytoplasma associated with the sugarcane white leaf (SCWL) disease. The experiment was conducted on plants of the sugarcane variety Khon Kaen 3 (KK3) infected with SCWL phytoplasma using insect vectors. Biochemical changes related to the SAR such as SA and total phenolic compounds were followed according to 4 different timepoints: 7, 14, 21 and 28 days after inoculation. Together, phytoplasma were quantified by RT-qPCR using the secA gene of phytoplasma. According to our results, the spraying of BTH and SA tended to increase the amounts of SA, total phenolic compounds and a lower presence of phytoplasma in the plants in comparison with the inoculated control. Spraying BTH at a concentration of 2.4 mM and SA at a concentration of 2.4 mM exhibited the best efficiency to reduce the concentration of phytoplasma. According to RT-qPCR results, the inoculated plants sprayed with BTH displayed a significantly lower concentration of phytoplasma compared to the inoculated controls. Overall, our results indicated that the spray of BTH and SA could induce an efficient SAR response to the phytoplasma associated with the SCWL disease. We expect these results will give support to the development of new products for controlling white leaf disease in sugarcane.

Differential expression between drought-tolerant and drought-sensitive sugarcane under mild and moderate water stress as revealed by a comparative analysis of leaf transcriptome.

Sugarcane contributes 80% of global sugar production and to bioethanol generation for the bioenergy industry. Its productivity is threatened by drought that can cause up to 60% yield loss. This study used RNA-Seq to gain a better understanding of the underlying mechanism by which drought-tolerant sugarcane copes with water stress. We compared gene expression in KPS01-12 (drought-tolerant genotype) and UT12 (drought-sensitive genotype) that have significantly different yield loss rates under drought conditions. We treated KPS01-12 and UT12 with mild and moderate water stress and found differentially expressed genes in various biological processes. KPS01-12 had higher expression of genes that were involved in water retention, antioxidant secondary metabolite biosynthesis, and oxidative and osmotic stress response than UT12. In contrast, the sensitive genotype had more down-regulated genes that were involved in photosynthesis, carbon fixation and Calvin cycle than the tolerant genotype. Our obtained expression profiles suggest that the tolerant sugarcane has a more effective genetic response than the sensitive genotype at the initiation of drought stress. The knowledge gained from this study may be applied in breeding programs to improve sugarcane production in drought conditions.

Outstanding Characteristics of Thai Non-GM Bred Waxy Cassava Starches Compared with Normal Cassava Starch, Waxy Cereal Starches and Stabilized Cassava Starches.

Waxy cassava roots of nine varieties successfully developed in Thailand by a non-genetic modification (non-GM), conventional breeding method were used for extracting starches and their starch physico-chemical properties were evaluated and compared with normal cassava starches, commercial waxy starches (i.e., waxy maize starch and waxy rice starch) and commercial stabilized starches (i.e., acetylated starch and hydroxypropylated starch). All waxy cassava varieties provided starches without amylose while normal cassava starches contained 18%-20% amylose contents. As determined by a Rapid Visco Analyzer (RVA) at 5% (dry basis), waxy cassava starches had the highest peak viscosity and the lowest setback viscosity. Cooked paste of waxy cassava starches had the greatest clarity and stability among all starches during storage at 4 ℃ for 7 days as evidenced by its high light transmittance (%T) at 650 nm. No syneresis was detected in waxy cassava starch gels after subjecting to four freeze-thaw cycles (4 weeks) indicating high potential use of waxy cassava starches, free from chemicals, to replace stabilized starches as thickening and texturing agents in food products.

Effect of pH on complex formation between debranched waxy rice starch and fatty acids.

Complex formations between debranched waxy rice starch (DBS) and fatty acids (FA) of different hydrocarbon chain lengths (8:0, 10:0, 12:0, 14:0, 16:0, and 18:0) were studied in an aqueous solution by measuring the blue colour stained with iodine. The objective of this study was to understand the effects of the solubility and hydrophobicity of guest molecules (FA) on the complex formation with DBS. Lauric acid (12:0) displayed the greatest complex forming ability with DBS by showing the least blue colour developed with iodine. The effect of pH (3-7) on the DBS/FA complex formation was evaluated by measuring the iodine-scanning spectra of the mixture. Short-chain FA (8:0) displayed less complex formation at pH>or=5, above the pK(a) of fatty acid (approximately 4.8), which suggested that the charge formation of the short-chain FA caused a lower partitioning of the FA into the hydrophobic cavity of the DBS single helix. On the contrary, FA of 10:0-18:0 displayed an increased complex formation at pH>5, which could be attributed to increased solubility of these longer-chain FA at a dissociated and ionized form. The hydrocarbon chain length of the FA had an important impact on the extent of the complex formation. A FA that had a shorter hydrocarbon chain was more soluble in an aqueous solution and more readily formed a complex with DBS. At pH 6 and 7 (above the pK(a)), 10:0 formed less inclusion complexes with DBS than did 12:0. Iodine-scanning spectra showed that the absorbances of all iodine-stained DBS/FA solutions at higher wavelength were substantially lower than that of the iodine-stained DBS alone, suggesting that FA preferentially formed inclusion complexes with DBS of longer chains.

Characterization of pectin extracted from banana peels of different varieties.

Pectins were extracted from banana peels of five different varieties using citric acid solution. The chemical characteristics of banana peel pectins were investigated and compared with citrus peel and apple pomace pectins which were extracted under the same extraction conditions to assess the potential of banana peels as an alternative source of commercial pectin. The yield of banana peel pectins ranged from 15.89 to 24.08%. The extracted banana peel pectins were categorized as high methoxyl pectin with the degree of esterification between 63.15 and 72.03% comparable to those of conventional pectin sources from citrus peel (62.83%) and apple pomace (58.44%). The anhydrouronic acid (AUA) content of banana peel pectins varied from 34.56 to 66.67%. Among various banana varieties being studied, pectin from Kluai Nam Wa variety had the highest AUA content (66.67%) which met the criteria for food additive pectin indicating its commercial significance as an alternative pectin source.

Structural and thermodynamic properties of rice starches with different genetic background Part 1. Differentiation of amylopectin and amylose defects.

A combined DSC - HPAEC-PAD approach, gel permeation chromatography and mild long-term acidic hydrolysis were employed to study the effects of amylopectin chain-length distributional and amylose defects on the assembly structures of amylopectin (crystalline lamellae, amylopectin clusters) in A-type polymorphic starches extracted from 11 Thai cultivars of rice with different amylose level. Joint analysis of the data allowed determining the contributions of different populations of amylopectin chains to the thermodynamic melting parameters of crystalline lamellae. It was shown that amylopectin chains with DP 6-12 and 25or=37 could be related to chains stabilizing these structures. The total effect of amylose and amylopectin defects can be described by means of Thomson-Gibbs' equation. The increase of defects in the assembly structures is accompanied by rise of the rates of acidic hydrolysis of both amorphous and crystalline parts in starches.

Influence of reaction parameters on carboxymethylation of rice starches with varying amylose contents.

The influence of reaction parameters on the carboxymethylation of rice starches with different amylose contents was investigated. Rice starches with varying amylose contents showed various degrees of susceptibility to the reaction conditions. The maximum degree of substitution (DS) for all three rice starches was obtained under similar reaction conditions which involved a reaction medium consisting of isopropanol-water at the ratio of 90:10, a molar ratio of NaOH:AGU at 1.5 and a reaction temperature and time of 40°C and 3 h. Under these conditions, the DS for all rice starches was similar; however, when the reaction was performed under conditions using lower NaOH concentration, the effect of starch types on the DS was observed. The results could be explained in terms of the granular/structural features of the different rice starches, their degrees of granular swelling as influenced by the reaction conditions and the accessibility of the etherifying reagents to starch molecules.

Glycolipid composition of Hevea brasiliensis latex.

Glycolipids of fresh latex from three clones of Hevea brasiliensis were characterized and quantified by HPLC/ESI-MS. Their fatty acyl and sterol components were further confirmed by GC/MS after saponification. The four detected glycolipid classes were steryl glucosides (SG), esterified steryl glucosides (ESG), monogalactosyl diacylglycerols (MGDG) and digalactosyl diacylglycerols (DGDG). Sterols in SG, ESG and total latex unsaponifiable were stigmasterol, ß-sitosterol and Δ5-avenasterol. The latter was found instead of fucosterol formerly described. Galactolipids were mainly DGDG and had a fatty acid composition different from that of plant leaves as they contained less than 5% C18:3. Glycolipids, which represented 27-37% of total lipids, displayed important clonal variations in the proportions of the different fatty acids. ESG, MGDG and DGDG from clone PB235 differed notably by their higher content in furan fatty acid, which accounted for more than 40% of total fatty acids. Clonal variation was also observed in the relative proportions of glycolipid classes except MGDG (8%), with 43-51% DGDG, 30-34% SG and 7-19% ESG. When compared with other plant cell content, the unusual glycolipid composition of H. brasiliensis latex may be linked to the peculiar nature of this specialized cytoplasm expelled from laticiferous system, especially in terms of functional and structural properties.

The fine structure of cassava starch amylopectin. Part 2: building block structure of clusters.

The aim of this work was to analyse the organization of unit chains inside clusters of cassava amylopectin. beta-Limit dextrins of the clusters and partly fragmented clusters (sub-clusters) were isolated previously [Laohaphatanaleart et al., Int. J. Biol. Macromol. (2010) doi:10.1016/j.ijbiomac.2010.01.0049] and were now hydrolysed extensively with the alpha-amylase (liquefying type) of Bacillus subtilis into small, branched building blocks. The blocks were size-fractionated and characterized structurally. The smallest blocks predominated in the clusters. They were single branched and possessed a degree of polymerization (DP) of 5-9. Blocks with DP 10-15 were double branched and constituted the second largest group. The clusters of cassava amylopectin, which were of rather uniform size, possessed typically 7-9 building blocks, and all clusters contained similar size-distributions of the blocks. The inter-block chain length was 7-8 residues. The possible mode of attack by the enzyme between the building blocks is discussed. A model of the building block organization in the clusters is presented, in which the structural roles of different sub-groups of clustered chains are suggested. A three-dimensional model suggests a possible organization of the building blocks inside the amorphous lamellae in the granular starch.

The fine structure of cassava starch amylopectin. Part 1: Organization of clusters.

The enzyme alpha-amylase from Bacillussubtilis was applied to partly hydrolyze purified cassava amylopectin into groups of clusters, which were called domains. The domains were further size-fractionated by methanol precipitation and then subjected to a second stage of alpha-amylolysis until the rate of hydrolysis was slow in order to release the single clusters. All domain and cluster fractions were hydrolyzed with beta-amylase into beta-limit dextrins. The size distribution and chain composition of the beta-limit dextrins were analyzed by gel-permeation chromatography and high-performance anion-exchange chromatography with pulsed amperometric detection, respectively. The sizes of the clusters in the form of beta-limit dextrins were uniform with an average degree of polymerization of 67-78. The distribution profiles of B-chains were similar in all cluster fractions, which suggested that the internal structure of the cassava amylopectin clusters was homogenous. Long B-chains were involved in the interconnection of clusters in the domain fractions. These were cleaved and a new group of chains of intermediate length was produced by the alpha-amylase together with short chains. In the isolated clusters, however, some chains corresponding to long B-chains still remained, which is not predicted by the traditional cluster model of the amylopectin structure. Instead, the alternative two-directional backbone model could explain the mode of interconnection between the clusters.

Application of bipolar electrodialysis on recovery of free lactic acid after simultaneous saccharification and fermentation of cassava starch.

The efficiency of bipolar electrodialysis (BED) for the recovery of lactic acid from fermentation broth was evaluated. Three systems of BED (bipolar-anion, bipolar-cation and bipolar-anion-cation) at fixed voltage (20 V) were compared using a model solution of ammonium lactate (100 g l(-1)). Results showed that bipolar-anion (BED-anion) was the most beneficial in terms of lactate flux, current efficiency, energy consumption and recovery ratio. Consequently, BED-anion was used to purify lactic acid from fermentation broth which had been pre-treated with mono-polar electrodialysis (MED). The final lactic acid concentration and lactate flux obtained were 144 g l(-1) and 393 g m(-2) h(-1), respectively. Using the two-step process (MED and BED-anion) the concentration of fermentation broth was increased by 33% and the total energy consumption was 2.76 kW h kg(-1).

Lactic acid production from sugar-cane juice by a newly isolated Lactobacillus sp.

A newly isolated sucrose-tolerant, lactic acid bacterium, Lactobacillus sp. strain FCP2, was grown on sugar-cane juice (125 g sucrose l(-1), 8 g glucose l(-1) and 6 g fructose l(-1)) for 5 days and produced 104 g lactic acid l(-1) with 90% yield. A higher yield (96%) and productivity (2.8 g l(-1 )h(-1)) were obtained when strain FCP2 was cultured on 3% w/v (25 g sucrose l(-1), 2 g glucose l(-1) and 1 g fructose l(-1)) sugar-cane juice for 10 h. Various cheap nitrogen sources such as silk worm larvae, beer yeast autolysate and shrimp wastes were also used as a substitute to yeast extract.