Japanese and Burmese have a vast way to address a specific person. Okell (1969) calls them as "pe... more Japanese and Burmese have a vast way to address a specific person. Okell (1969) calls them as "personal referents" in his Burmese description. This notion can be adapted to Japanese. Personal referents include personal pronouns, person's names, kin terms, job titles and so on. A speaker must choose an appropriate noun from these, based on a given situation. In addition, the two languages share the honorifics, which are added to a person's name to express the speaker's attitude to the hearer. According to these linguistic phenomena, it is necessary to know features that each personal referent has and to clarify the criteria or conditions to choose an appropriate one, especially when establishing an automatic translation system. This paper aims to describe some basic features of Burmese personal referents to contribute construction of an algorithm to get the right output theoretically.
Many languages in East and Southeast Asia exhibit an intensive use of pronoun substitutes, such a... more Many languages in East and Southeast Asia exhibit an intensive use of pronoun substitutes, such as kinship terms, that refer to speakers and addressees in the way that personal pronouns do although they are not considered personal pronouns. The distribution of pronoun substitutes overlaps substantially with that of address terms. This study thus provides an overview of previous studies on pronoun substitutes and address terms in Thai, Burmese, Malay, Indonesian, Javanese and Korean and covers the following issues: whether the two phenomena are distinguished, the terminology used for them, their formal and semantic characteristics, their relation to personal pronouns and how pronoun substitutes differ from the so-called "imposters" (Collins & Postal 2012) in English.
It is widely known that many lexical verbs are also used as functional words in Burmese. Used as ... more It is widely known that many lexical verbs are also used as functional words in Burmese. Used as functional words, some of these verbs precede the main verb, and some fo llow. A fe w verbs can both precede and fo llow the main verb. For example, when preceding the main verb, the verb 6� tGi 'look' has an adverbial function meaning something like 'with discretion, carefully'; when it fo llows the head-verb, it means 'try to [verb] '. The verb GO: pe 'give' also has functional usages. Following the head verb, its function is benefactive (or applicative), as in (1), or else it denotes the 'destination of action', as in (2) (the opposite direction, with UK ju 'take' is shown in (3)). It can also have a transitive/causative meaning, as in (4).) (' (' (' ('/ / 0 (' (' (1 oaJ;GOXl c;>c'1P:3dO'6m 3da>0: or (9O?GO:�OOIl tGano khlNbja.7atwd/?asci/ go pha? pe.& 1 you.for/on behalf of/to read.pe.lRR 'I will read it fo r/on behalf of/to you.' () 0 (' & 0 (' 2 �. 3dG(9 O?<J?O?oo'P 3dOOOUa>0 8lJ.7aphe tdka80.l1Ja ?INgalei? sa his.father university.in English.language 'His fa ther teaches English at university. ' Q 0 C" & 0 C' (3) O(. O)�: O?<J?O?oo'P 3dOOOUa>0 8lJ.7aphe tdka80.l1Ja ?INgalei? sa (' (' O)CGU:O?OOIl elN.pe.& learn.give.REAL (' (' o)CO(O?OOIl 81NJu.d£ his.daughter university.in English.language 'His daughter learns English at university.' learn. take. REAL
In order to achieve direct fermentation of an optically pure d-lactic acid from cellulosic materi... more In order to achieve direct fermentation of an optically pure d-lactic acid from cellulosic materials, an endoglucanase from a Clostridium thermocellum (CelA)-secreting plasmid was introduced into an l-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum (∆ldhL1) bacterial strain. CelA expression and its degradation of β-glucan was confirmed by western blot analysis and enzyme assay, respectively. Although the CelA-secreting ∆ldhL1 assimilated cellooligosaccharides up to cellohexaose (although not cellotetraose), the main end product was acetic acid, not lactic acid, due to the conversion of lactic acid to acetic acid. Cultivation under anaerobic conditions partially suppressed this conversion resulting in the production of 1.27 g/l of D-lactic acid with a high optical purity of 99.5% from a medium containing 2 g/l of cellohexaose. Subsequently, D-lactic acid fermentation from barley β-glucan was carried out with the addition of Aspergillus aculeatus β-glucosidase produced by recombinant Aspergillus oryzae and 1.47 g/l of D-lactic was produced with a high optical purity of 99.7%. This is the first report of direct lactic acid fermentation from β-glucan and a cellooligosaccharide that is a more highly polymerized sugar than cellotriose.
as the pressure media. Moreover, the production of HC with could be further improved by the addit... more as the pressure media. Moreover, the production of HC with could be further improved by the addition of H 2 O 2 (60 mmol/L) and the relative yield of HC in moderate pressure was enhanced by over 4.5k in comparison with the control. This study offered a new approach for improving the industrial production of HC.
In order to achieve efficient d-lactic acid fermentation from a mixture of xylose and glucose, th... more In order to achieve efficient d-lactic acid fermentation from a mixture of xylose and glucose, the xylose-assimilating xylAB operon from Lactobacillus pentosus (PXylAB) was introduced into an l-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum (ΔldhL1-xpk1::tkt-Δxpk2) strain in which the phosphoketolase 1 gene (xpk1) was replaced with the transketolase gene (tkt) from Lactococcus lactis, and the phosphoketolase 2 (xpk2) gene was deleted. Two copies of xylAB introduced into the genome significantly improved the xylose fermentation ability, raising it to the same level as that of ΔldhL1-xpk1::tkt-Δxpk2 harboring a xylAB operon-expressing plasmid. Using the two-copy xylAB integrated strain, successful homo-d-lactic acid production was achieved from a mixture of 25 g/l xylose and 75 g/l glucose without carbon catabolite repression. After 36-h cultivation, 74.2 g/l of lactic acid was produced with a high yield (0.78 g per gram of consumed sugar) and an optical purity of d-lactic acid of 99.5%. Finally, we successfully demonstrated homo-d-lactic acid fermentation from a mixture of three kinds of sugar: glucose, xylose, and arabinose. This is the first report that describes homo-d-lactic acid fermentation from mixed sugars without carbon catabolite repression using the xylose-assimilating pathway integrated into lactic acid bacteria.
In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw co... more In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw corn starch, we constructed L-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum and introduced a plasmid encoding Streptococcus bovis 148 ␣-amylase (AmyA). The resulting strain produced only D-lactic acid from glucose and successfully expressed amyA. With the aid of secreting AmyA, direct D-lactic acid fermentation from raw corn starch was accomplished. After 48 h of fermentation, 73.2 g/liter of lactic acid was produced with a high yield (0.85 g per g of consumed sugar) and an optical purity of 99.6%. Moreover, a strain replacing the ldhL1 gene with an amyA-secreting expression cassette was constructed. Using this strain, direct D-lactic acid fermentation from raw corn starch was accomplished in the absence of selective pressure by antibiotics. This is the first report of direct D-lactic acid fermentation from raw starch.
In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw co... more In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw corn starch, we constructed L-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum and introduced a-amylase from a Streptococcus bovis 148 (AmyA)-secreting plasmid. The resulting strain produced only D-lactic acid from glucose and successfully expressed amyA. With the aid of secreting AmyA, direct D-lactic acid fermentation from raw corn starch was accomplished. After 48 h of fermentation, 73.4 g/l of lactic acid was produced with high yield (0.85 g/g of consumed sugar) and an optical purity of 99.6k. Moreover, a strain substituting the ldhL1 gene with an amyA-secreting expression cassette was constructed. Using this strain, direct D-lactic acid fermentation from raw corn starch was accomplished in the absence of selective pressure by antibiotics.
We developed a new cell surface engineering system based on the PgsA anchor protein from Bacillus... more We developed a new cell surface engineering system based on the PgsA anchor protein from Bacillus subtilis. In this system, the N terminus of the target protein was fused to the PgsA protein and the resulting fusion protein was expressed on the cell surface. Using this new system, we constructed a novel starch-degrading strain of Lactobacillus casei by genetically displaying ␣-amylase from the Streptococcus bovis strain 148 with a FLAG peptide tag (AmyAF). Localization of the PgsA-AmyA-FLAG fusion protein on the cell surface was confirmed by immunofluorescence microscopy and flow cytometric analysis. The lactic acid bacteria which displayed AmyAF showed significantly elevated hydrolytic activity toward soluble starch. By fermentation using AmyAF-displaying L. casei cells, 50 g/liter of soluble starch was reduced to 13.7 g/liter, and 21.8 g/liter of lactic acid was produced within about 24 h. The yield in terms of grams of lactic acid produced per gram of carbohydrate utilized was 0.60 g per g of carbohydrate consumed at 24 h. Since AmyA was immobilized on the cells, cells were recovered after fermentation and used repeatedly. During repeated utilization of cells, the lactic acid yield was improved to 0.81 g per g of carbohydrate consumed at 72 h. These results indicate that efficient simultaneous saccharification and fermentation from soluble starch to lactic acid were carried out by recombinant L. casei cells with cell surface display of AmyA.
The 5′-untranslated leader sequence (UTLS) of the slpA gene from Lactobacillus acidophilus contri... more The 5′-untranslated leader sequence (UTLS) of the slpA gene from Lactobacillus acidophilus contributes to mRNA stabilization by producing a 5′ stem and loop structure, and a high-level expression system for the lactic acid bacteria was developed using the UTLS in this study. A plasmid, which expresses α-amylase under the control of the ldh promoter, was constructed by integrating the core promoter sequence with the UTLS. The role of the UTLS in increasing the copies of the α-amylase mRNA was proved by measuring α-amylase activity in the culture supernatant and the relative expression of α-amylase mRNA was determined by the quantitative real-time PCR analysis. Moreover, several expression systems were constructed by combining the core promoter sequence with the UTLS or with the partially deleted UTLS and the expression level was evaluated. The use of the UTLS led to the success in improving α-amylase expression in the two strains of Lactobacillus casei and Lactococcus lactis. The current study showed that the improvement in protein production using the UTLS could be applied to the expression system in the lactic acid bacteria.
We have developed a novel Escherichia coli cell surface display system by employing PgsA as an an... more We have developed a novel Escherichia coli cell surface display system by employing PgsA as an anchoring motif. In our display system, C-terminal fusion to PgsA anchor protein from Bacillus subtilis was used. The enzymes selected for display were α-amylase (AmyA) from Streptococcus bovis 148 and lipase B (CALB) from Candida antarctica. The molecular mass values of AmyA and CALB are approximately 77 and 34 kDa, respectively. The enzymes were displayed on the surface as a fusion protein with a FLAG peptide tag at the C terminus. Both the PgsA-AmyA-FLAG and PgsA-CALB-FLAG fusion proteins were shown to be displayed by immunofluorescence labeling using anti-FLAG antibody. The displayed enzymes were active forms, and AmyA and CALB activities reached 990 U/g (dry cell weight) and 4.6 U/g (dry cell weight), respectively. AmyA-displaying E. coli cells grew utilizing cornstarch as the sole carbon source, while CALB-displaying E. coli cells catalyzed enantioselective transesterification, indicating that they are effective whole-cell biocatalysts. Since a target enzyme with a size of 77 kDa and an industrially useful lipase have been successfully displayed on the cell surface of E. coli for the first time, PgsA protein is probably a useful anchoring motif to display various enzymes.
Japanese and Burmese have a vast way to address a specific person. Okell (1969) calls them as "pe... more Japanese and Burmese have a vast way to address a specific person. Okell (1969) calls them as "personal referents" in his Burmese description. This notion can be adapted to Japanese. Personal referents include personal pronouns, person's names, kin terms, job titles and so on. A speaker must choose an appropriate noun from these, based on a given situation. In addition, the two languages share the honorifics, which are added to a person's name to express the speaker's attitude to the hearer. According to these linguistic phenomena, it is necessary to know features that each personal referent has and to clarify the criteria or conditions to choose an appropriate one, especially when establishing an automatic translation system. This paper aims to describe some basic features of Burmese personal referents to contribute construction of an algorithm to get the right output theoretically.
Many languages in East and Southeast Asia exhibit an intensive use of pronoun substitutes, such a... more Many languages in East and Southeast Asia exhibit an intensive use of pronoun substitutes, such as kinship terms, that refer to speakers and addressees in the way that personal pronouns do although they are not considered personal pronouns. The distribution of pronoun substitutes overlaps substantially with that of address terms. This study thus provides an overview of previous studies on pronoun substitutes and address terms in Thai, Burmese, Malay, Indonesian, Javanese and Korean and covers the following issues: whether the two phenomena are distinguished, the terminology used for them, their formal and semantic characteristics, their relation to personal pronouns and how pronoun substitutes differ from the so-called "imposters" (Collins & Postal 2012) in English.
It is widely known that many lexical verbs are also used as functional words in Burmese. Used as ... more It is widely known that many lexical verbs are also used as functional words in Burmese. Used as functional words, some of these verbs precede the main verb, and some fo llow. A fe w verbs can both precede and fo llow the main verb. For example, when preceding the main verb, the verb 6� tGi 'look' has an adverbial function meaning something like 'with discretion, carefully'; when it fo llows the head-verb, it means 'try to [verb] '. The verb GO: pe 'give' also has functional usages. Following the head verb, its function is benefactive (or applicative), as in (1), or else it denotes the 'destination of action', as in (2) (the opposite direction, with UK ju 'take' is shown in (3)). It can also have a transitive/causative meaning, as in (4).) (' (' (' ('/ / 0 (' (' (1 oaJ;GOXl c;>c'1P:3dO'6m 3da>0: or (9O?GO:�OOIl tGano khlNbja.7atwd/?asci/ go pha? pe.& 1 you.for/on behalf of/to read.pe.lRR 'I will read it fo r/on behalf of/to you.' () 0 (' & 0 (' 2 �. 3dG(9 O?<J?O?oo'P 3dOOOUa>0 8lJ.7aphe tdka80.l1Ja ?INgalei? sa his.father university.in English.language 'His fa ther teaches English at university. ' Q 0 C" & 0 C' (3) O(. O)�: O?<J?O?oo'P 3dOOOUa>0 8lJ.7aphe tdka80.l1Ja ?INgalei? sa (' (' O)CGU:O?OOIl elN.pe.& learn.give.REAL (' (' o)CO(O?OOIl 81NJu.d£ his.daughter university.in English.language 'His daughter learns English at university.' learn. take. REAL
In order to achieve direct fermentation of an optically pure d-lactic acid from cellulosic materi... more In order to achieve direct fermentation of an optically pure d-lactic acid from cellulosic materials, an endoglucanase from a Clostridium thermocellum (CelA)-secreting plasmid was introduced into an l-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum (∆ldhL1) bacterial strain. CelA expression and its degradation of β-glucan was confirmed by western blot analysis and enzyme assay, respectively. Although the CelA-secreting ∆ldhL1 assimilated cellooligosaccharides up to cellohexaose (although not cellotetraose), the main end product was acetic acid, not lactic acid, due to the conversion of lactic acid to acetic acid. Cultivation under anaerobic conditions partially suppressed this conversion resulting in the production of 1.27 g/l of D-lactic acid with a high optical purity of 99.5% from a medium containing 2 g/l of cellohexaose. Subsequently, D-lactic acid fermentation from barley β-glucan was carried out with the addition of Aspergillus aculeatus β-glucosidase produced by recombinant Aspergillus oryzae and 1.47 g/l of D-lactic was produced with a high optical purity of 99.7%. This is the first report of direct lactic acid fermentation from β-glucan and a cellooligosaccharide that is a more highly polymerized sugar than cellotriose.
as the pressure media. Moreover, the production of HC with could be further improved by the addit... more as the pressure media. Moreover, the production of HC with could be further improved by the addition of H 2 O 2 (60 mmol/L) and the relative yield of HC in moderate pressure was enhanced by over 4.5k in comparison with the control. This study offered a new approach for improving the industrial production of HC.
In order to achieve efficient d-lactic acid fermentation from a mixture of xylose and glucose, th... more In order to achieve efficient d-lactic acid fermentation from a mixture of xylose and glucose, the xylose-assimilating xylAB operon from Lactobacillus pentosus (PXylAB) was introduced into an l-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum (ΔldhL1-xpk1::tkt-Δxpk2) strain in which the phosphoketolase 1 gene (xpk1) was replaced with the transketolase gene (tkt) from Lactococcus lactis, and the phosphoketolase 2 (xpk2) gene was deleted. Two copies of xylAB introduced into the genome significantly improved the xylose fermentation ability, raising it to the same level as that of ΔldhL1-xpk1::tkt-Δxpk2 harboring a xylAB operon-expressing plasmid. Using the two-copy xylAB integrated strain, successful homo-d-lactic acid production was achieved from a mixture of 25 g/l xylose and 75 g/l glucose without carbon catabolite repression. After 36-h cultivation, 74.2 g/l of lactic acid was produced with a high yield (0.78 g per gram of consumed sugar) and an optical purity of d-lactic acid of 99.5%. Finally, we successfully demonstrated homo-d-lactic acid fermentation from a mixture of three kinds of sugar: glucose, xylose, and arabinose. This is the first report that describes homo-d-lactic acid fermentation from mixed sugars without carbon catabolite repression using the xylose-assimilating pathway integrated into lactic acid bacteria.
In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw co... more In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw corn starch, we constructed L-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum and introduced a plasmid encoding Streptococcus bovis 148 ␣-amylase (AmyA). The resulting strain produced only D-lactic acid from glucose and successfully expressed amyA. With the aid of secreting AmyA, direct D-lactic acid fermentation from raw corn starch was accomplished. After 48 h of fermentation, 73.2 g/liter of lactic acid was produced with a high yield (0.85 g per g of consumed sugar) and an optical purity of 99.6%. Moreover, a strain replacing the ldhL1 gene with an amyA-secreting expression cassette was constructed. Using this strain, direct D-lactic acid fermentation from raw corn starch was accomplished in the absence of selective pressure by antibiotics. This is the first report of direct D-lactic acid fermentation from raw starch.
In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw co... more In order to achieve direct and efficient fermentation of optically pure D-lactic acid from raw corn starch, we constructed L-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum and introduced a-amylase from a Streptococcus bovis 148 (AmyA)-secreting plasmid. The resulting strain produced only D-lactic acid from glucose and successfully expressed amyA. With the aid of secreting AmyA, direct D-lactic acid fermentation from raw corn starch was accomplished. After 48 h of fermentation, 73.4 g/l of lactic acid was produced with high yield (0.85 g/g of consumed sugar) and an optical purity of 99.6k. Moreover, a strain substituting the ldhL1 gene with an amyA-secreting expression cassette was constructed. Using this strain, direct D-lactic acid fermentation from raw corn starch was accomplished in the absence of selective pressure by antibiotics.
We developed a new cell surface engineering system based on the PgsA anchor protein from Bacillus... more We developed a new cell surface engineering system based on the PgsA anchor protein from Bacillus subtilis. In this system, the N terminus of the target protein was fused to the PgsA protein and the resulting fusion protein was expressed on the cell surface. Using this new system, we constructed a novel starch-degrading strain of Lactobacillus casei by genetically displaying ␣-amylase from the Streptococcus bovis strain 148 with a FLAG peptide tag (AmyAF). Localization of the PgsA-AmyA-FLAG fusion protein on the cell surface was confirmed by immunofluorescence microscopy and flow cytometric analysis. The lactic acid bacteria which displayed AmyAF showed significantly elevated hydrolytic activity toward soluble starch. By fermentation using AmyAF-displaying L. casei cells, 50 g/liter of soluble starch was reduced to 13.7 g/liter, and 21.8 g/liter of lactic acid was produced within about 24 h. The yield in terms of grams of lactic acid produced per gram of carbohydrate utilized was 0.60 g per g of carbohydrate consumed at 24 h. Since AmyA was immobilized on the cells, cells were recovered after fermentation and used repeatedly. During repeated utilization of cells, the lactic acid yield was improved to 0.81 g per g of carbohydrate consumed at 72 h. These results indicate that efficient simultaneous saccharification and fermentation from soluble starch to lactic acid were carried out by recombinant L. casei cells with cell surface display of AmyA.
The 5′-untranslated leader sequence (UTLS) of the slpA gene from Lactobacillus acidophilus contri... more The 5′-untranslated leader sequence (UTLS) of the slpA gene from Lactobacillus acidophilus contributes to mRNA stabilization by producing a 5′ stem and loop structure, and a high-level expression system for the lactic acid bacteria was developed using the UTLS in this study. A plasmid, which expresses α-amylase under the control of the ldh promoter, was constructed by integrating the core promoter sequence with the UTLS. The role of the UTLS in increasing the copies of the α-amylase mRNA was proved by measuring α-amylase activity in the culture supernatant and the relative expression of α-amylase mRNA was determined by the quantitative real-time PCR analysis. Moreover, several expression systems were constructed by combining the core promoter sequence with the UTLS or with the partially deleted UTLS and the expression level was evaluated. The use of the UTLS led to the success in improving α-amylase expression in the two strains of Lactobacillus casei and Lactococcus lactis. The current study showed that the improvement in protein production using the UTLS could be applied to the expression system in the lactic acid bacteria.
We have developed a novel Escherichia coli cell surface display system by employing PgsA as an an... more We have developed a novel Escherichia coli cell surface display system by employing PgsA as an anchoring motif. In our display system, C-terminal fusion to PgsA anchor protein from Bacillus subtilis was used. The enzymes selected for display were α-amylase (AmyA) from Streptococcus bovis 148 and lipase B (CALB) from Candida antarctica. The molecular mass values of AmyA and CALB are approximately 77 and 34 kDa, respectively. The enzymes were displayed on the surface as a fusion protein with a FLAG peptide tag at the C terminus. Both the PgsA-AmyA-FLAG and PgsA-CALB-FLAG fusion proteins were shown to be displayed by immunofluorescence labeling using anti-FLAG antibody. The displayed enzymes were active forms, and AmyA and CALB activities reached 990 U/g (dry cell weight) and 4.6 U/g (dry cell weight), respectively. AmyA-displaying E. coli cells grew utilizing cornstarch as the sole carbon source, while CALB-displaying E. coli cells catalyzed enantioselective transesterification, indicating that they are effective whole-cell biocatalysts. Since a target enzyme with a size of 77 kDa and an industrially useful lipase have been successfully displayed on the cell surface of E. coli for the first time, PgsA protein is probably a useful anchoring motif to display various enzymes.
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Papers by Kenji OKANO